Nonsteroidal and steroidal compounds with potent androgen receptor down-regulation and anti prostate cancer activity

ABSTRACT

Nonsteroid and steroid compounds that cause down-regulation of the androgen receptor (AR), both full length and splice variant, induce apoptosis and inhibit proliferation of inhibiting proliferation and migration of androgen sensitive cancer cells. The steroid compounds and nonsteroid compounds may be agents for the prevention and/or treatment of cancer, including prostate cancer, castration resistant prostate cancer, bladder cancer, pancreatic cancer, hepatocellular carcinoma, benign prostatic hyperplasia (BPH), Kennedy&#39;s disease, androgenetic alopecia, breast cancer, androgen-insensitive syndrome, and spinal and bulbar muscular atrophy.

CROSS REFERENCE

This application claims benefit of U.S. Provisional Application No.61/808,345, filed on Apr. 4, 2013, and U.S. Provisional Application No.61/808,902, filed on Apr. 5, 2013, both which are herein incorporated byreference.

STATEMENT OF FEDERALLY SPONSORED RESEARCH

This invention was made with government support under grant numberCA129379 awarded by the National Institutes of Health. The governmenthas certain rights in the invention.

BACKGROUND OF THE INVENTION

Prostate cancer (PC) is the most common tumor in men in westerncountries, and the second cause of cancer-related death among them.Almost 80% of cases are diagnosed as localized disease, and radiation orsurgery can be curative. However, despite current treatment options,there is still a relapse rate of 30-60%.

Applicant has designed and synthesized of novel nonsteroid andsteroid-mimetic compounds that cause down-regulation of the androgenreceptor (AR), both full length and splice variant, induce apoptosis andinhibit proliferation of both androgen sensitive and castrationresistant human prostate cancer cells. These steroid compounds andnonsteroid compounds may be agents for the prevention and/or treatmentof all forms of prostate cancer and all other diseases that depend onfunctional AR.

AR is a well established target for therapeutic intervention in prostatecancer.

Other androgen receptor associated conditions that may be preventedand/or treated in the present invention include bladder cancer,pancreatic cancer, hepatocellular carcinoma, benign prostatichyperplasia (BPH) and Kennedy's disease.

Androgen and androgen receptor (AR) play crucial role in the developmentand advancement of PC. As a consequence, for locally advanced ormetastatic disease, hormonal treatment with androgen deprivationtherapy, which blocks the production (CYP17 inhibitors: Abiraterone; 1)and/or activity of androgen (anti-androgens, Bicalutamide; 2), is astandard approach for the majority of patients (Chart 1), but most casesthe duration of response is limited to 12-24 months, and the diseasewill become castration-resistant (CRPC) with no treatment options.Approximately 85% of CRPC patients succumb within 5 years and docetaxel(3) is currently the only treatment shown to provide even minimalsurvival benefit.

In castration-resistant environment aberrant AR reactivation isimplicated through numerous mechanisms which leads to over expression ofmutated AR, AR amplification and local androgen synthesis.⁵ Recently,multiple alternative spliced AR isoforms (AR-Vs) have been identified inCRPC. Unlike full-length AR (fAR) these AR-Vs are lack of thehormone-binding domain and also their activities are independent ofhormone. The AR-Vs are expressed at higher level in various tumors andare three to five times more potent than fAR in transactivatingactivity. Patients expressing constitutively active AR-Vs will in thelong-term probably not benefit from antiandrogen and therapies aiming toreduce androgen synthesis. In fact, ‘not all CRPC patients do howeverrespond to novel antiandrogen (MDV3100; 4) and CYP17 inhibitors (1), andeven those who do subsequently relapse within a few months’. Based onabove findings it is envisioned that effective treatment of CRPCpatients will require new drugs that can modulate all forms of AR suchas AR down-regulating (ARD) agents (ARDAs). The substantial anti-PCefficacy of Phase II clinical candidate ‘Galetereone’ (VN/124-1, 5,Chart 1 below) in comparison to abiraterone is by virtue of its multitarget mechanism of action (CYP-17 inhibition, antiandrogen and ARDactivity). (http://clinicaltrials.gov/ct2/show/NCT01709734).

For the development of ARD agents Applicant recently systematicallyexplored the structure of 5 (VN/124-1). In which chemical functionsatC17, C16 and C3 of Galeterone modified while androstene scaffoldunaltered. The C17 bezimidazole (BzIm) ring in Galeterone is essentialto block catalytic 17α-hydroxylation of pregnenolone and progesteronemediated by CYP17 enzyme to obtain bio-precursors of androgenbiosynthesis. Where 3β-OH group forms single direct hydrogen bondbetween inhibitor and polar amino acid residue of CYP17 enzyme.Similarly 3β-OH or 3-keto group of natural and synthetic hormones areidentical of the interactions that are conserved in the androgen,estrogen, glucocorticoid, mineralocorticoid and progesterone receptors.Therefore, H-bonding interaction of 3β-OH and keto group of syntheticand natural hormones are critical for ligand recognition by CYP17 andhormone receptors. The 3β-OH group in galeterone is responsible forpotent CYP17 and antiandrogen activity. This is further supported byloss of antiandrogen and CYP17 inhibitory activity of galeterone onmodification of its C3 substitution with imidazole carabamate (VNPT55(6)) and pyridine carboxylate (VNPT178 (7)) group. This modificationsalso enhanced ARD by 8.25-fold against fAR, 4-fold against AR-Vs and4-fold increase in anti-PC activity. Applicant has successfullydiscovered and reported first rather selective (Chart 2) ARdown-regulating agents (both fAR and AR-Vs).

Chart 1: Chemical Structure of Compounds 1-5 (Clinical Anti PC Agents).

Chart 2: Chemical Structure of Selective ARD Agents

BRIEF SUMMARY OF THE INVENTION

Considering the of AR modulating strategy (ARdown-regulation/degradation) in the treatment of prostate cancer diseaseat present Applicant has designed, synthesized and tested several novelnonsteroidal and steroidal small molecules as antiprostate canceragents. A preliminary account of part of this work has recently beenreported.²¹ The advantages may include improved physiochemical &pharmacokinetic properties, increased selectivity of biological activityand, as drug resistance remain a major problem in cancer therapy, thevariety of well-tolerated ARDAs with differing chemical structures offerthe possibility that several useful drugs will be available.

For the design of novel nonsteroidal agents Applicant has replacedandrostene core with some reported steroidal surrogates such asbiphenyl,^(22, 23) naphthalene,²² stilbene,²⁴ tetrahydroisoquinoline anddesigned diaryl compound with alkylamine, amide and sulfonamide linkerscore structure.²⁵ On all these scaffolds head group (BzIm) is heldconstant while tail is hydroxyl or masked with methyl or replaced withcarboxylic/nitrile/imidazole/tetrazole etc groups in combination.Similarly for steroidal agents Applicant chose androstene and estrogenstructure as scaffolds in which bezimidazole as head and variousfunction such as acid, ester, imidazole, ethers carbamates etc functionsas tail.

Biological Activity

Concept of AR Down-Regulation as a Therapeutic Strategy

Current treatments are centered on blocking androgen-signaling axis, butthe emergence of castration resistant PC (CRCP) prevails. Such resistantPC tumors continue to grow in the presence of low circulating endogenousligands by virtue of the presence of active and functional AR. Applicanthas developed novel AR down-regulating agents (ARDAs) to treat and/orprevent cancer.

Previously Applicant discovered few ARDAs (synthetic small molecules)through HipHop Pharmacophore modeling.¹

Applicant has developed new steroidal derivatives as potent ARDA vialead optimization of our PC drug candidate, VN/124-1 (Galeterone; FIG.1), currently in phase II clinical trials.²

The instant invention includes novel nonsteroidal compounds and novelsteroid compounds as ARDAs.

The instant invention includes a method of treating prostate cancercomprising administering an effective amount of the compounds of theinstant invention.

The instant invention includes a method of treating castration resistantprostate cancer.

The instant invention includes degrading a full length or a splicevariant by administering an effective amount of the compounds of theinstant invention.

The instant invention includes a method of inhibiting proliferation ofandrogen sensitive cells by administering an effective amount of thecompounds of the instant invention.

The compounds may be used for treatment individually or with one or morecompounds of the instant invention. The compounds may also be used fortreatment with other known compounds or treatment methods.

C-17 Benzimidazole (BzIm) is essential and optimal for observed androgenreceptor down-regulating (ARD) activity. Replacement of 3β-OH withimidazolecarbamate (FIG. 1) resulted in potent and specific ARDactivity. The steroidal molecular frame of VN/124-1 was not altered inprevious study.

Although side effects for VN/124-1 due to its steroidal scaffold havenot been observed in the clinic thus far, the potential side effects ofsteroidal drugs believed to be due to binding to various steroidreceptors to elicit agonistic or antagonistic effects may be a matter ofconcern.

In the last two decades several reports have focused on diverse chemicalscaffolds such as biphenyl, naphthylene, and stibene, as excellentsurrogates for the steroidal scaffold in the breast and prostate cancersdrug discovery and development fields.

The present invention includes possible modification of steroidalscaffold while retaining C-17 BzIm group and retaining or replacing C-3hydroxyl group with hydrophilic moieties, including OCH₃, COOH, CN, and1H-tetrazole groups.

Agents discovered by Applicant include selected biphenyl,naphthylbipheyl and stilbene scaffolds (FIG. 2). Applicant's inventionincludes Flexible alignment experiment using (MOE) for nonsteroidalscaffolds to find out how well hydrophilic and hydrophobic functions onnew scaffolds superimposed with VN/124-1 (FIG. 3). In addition, theestrone derivatives were designed as alternative of the androstane coreof our earlier compounds.

DESCRIPTION OF THE DRAWINGS

FIG. 1. Lead optimization of VN/124-1.

FIG. 2. Novel steroid surrogates.

FIG. 3. Flexible alignment with VN/124-1.

FIG. 4. Effects of Compounds on Transcriptional Activity of LuciferaseMediated Via LNCaP-AR in LNCaP-lu Prostate Cancer Cells.

FIG. 5. Antiproliferative activity against androgen responsive LNCaPcells (A), CRPC cell line C4-2B (B) by MTT assay.

FIG. 6. Antiproliferative activity against androgen responsive cell lineCRPC cell line C4-2B by MTT assay.

FIG. 7. Screening of new compounds (compared to VN/124-1) for inhibitionof cell growth @ 1 and 10 μM using MTT assay.

FIG. 8. Effect of compounds on suppression of AR protein levels in LNCaPcells (15 μM of each compound for 24 h.

FIG. 9A-H. Effects of compounds on oncogenes.

FIG. 10. Effects of VN124-1 and analogs on cell migration.

FIG. 11. Example of HPLC chromatogram of three component mixture. Theretention times for VNPP433-6β, VNPP433-3α(A3) and VNPP433-3β(2d)(A1)were 5.14, 7.87 and 10.07 min, respectively.

FIG. 12. Mechanisms of induced AR degradation in prostate cancer cells

FIG. 13. Therapeutic approaches to block androgen receptor (AR)transactivation

FIG. 14. Diagram of androgen receptor protein

DETAIL DESCRIPTION OF THE INVENTION

Synthesis

Synthetic methods of molecules in this study are depicted in the Schemesbelow. The key intermediates, alkylated-BzIm's (1a, 2a) synthesized byfollowing reported KOH in DMSO method, while arylated-BzIm (11a) byreported method of ligated-catalysis' Ullmann condensation reaction.Suzuki coupling of bromoalkyl/aryl-BzIm with corresponding boronic acidresulted in biphenyls (1-8, 11). Conversion of nitrile-biphenyls (3, 4)to 1H-tetrazoles (9, 10) was achieved by 1,3-dipolar addition of sodiumazide in presence of ammonium chloride in DMF. Styrenes (12a,13a) wereobtained by witting method¹⁰ subjected to Mizoroki-Heck reaction to getdesired (E)-stilbenes (12, 13). For estrone-3-carboxylate (16), 3-enolcarbon was activated by triflate followed by replacement withcarboxylate through pd catalyzed reaction. BzIm substitution on estroneC-17 via Vilsmeir reaction achieved following our routine method for thesynthesis of C-17 BzIm substituted steroid compounds. The 17-pyridylestrone derivative synthesized following reported procedure.¹³

See also the schemes below.Novel Nonsteroids for Cancer Treatment:Biphenyl Derivatives (B Series)

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —N(R′)₂—, —SH, —OMe, —CN, —COON, —COOMe,—COOEt, imidazole, 1H-tetrazole and their combinations, wherein each R′is independently selected from alkyl or aryl.Examples of B series

-   -   n=0 or 1 or 2

TABLE 1 Biphenyl derivatives (B series) VNPP347 (B1)3-[4-(benzimidazolylmethyl)phenyl]-1-methoxybenzene VNPP358 (B2)3-[4-(benzimidazolylmethyl)phenyl]benzenecarbonitrile VNPP356(B3)3-[4-(benzimidazolylmethyl)phenyl]phenol B44-[4-(benzimidazolylmethyl)phenyl]-2-methoxybenzenecarbonitrile B54-[4-(benzimidazolylmethyl)phenyl]-2-methoxyphenol B65-[4-(benzimidazolylmethyl)phenyl]-2-hydroxybenzenecarbonitrile VNPP360(B7) 3-[4-(benzimidazolylmethyl)phenyl]benzoic acid B85-[4-(benzimidazolylmethyl)phenyl]-2-methoxybenzoic acid B94-[4-(benzimidazolylmethyl)phenyl]-1,2-dimethoxybenzene B105-[4-(benzimidazolylmethyl)phenyl]-2-methoxybenzenecarbonitrile VNPP361(B11) 5-{3-[4-(benzimidazolylmethyl)phenyl]phenyl}-1H-1,2,3,4-tetraazoleB12 2-(1H-1,2,3,4-tetraazol-5-yl)-4-[4-(benzimidazolylmethyl)phenyl]-1-methoxybenzene B132-(1H-1,2,3,4-tetraazol-5-yl)-4-[4-(benzimidazolylmethyl)phenyl]phenolB14 {[4-(3-imidazolylphenyl)phenyl]methyl}benzimidazole B154-[4-(benzimidazolylmethyl)phenyl]-2-imidazolyl-1-methoxybenzene B164-[4-(benzimidazolylmethyl)phenyl]-2-imidazolylbenzoic acid B174-[4-(benzimidazolylmethyl)phenyl]-2-imidazolylbenzenecarbonitrileVNPP321 (B18) 3-[4-(2-benzimidazolylethyl)phenyl]-1-methoxybenzeneVNPP355(B19) 3-[4-(2-benzimidazolylethyl)phenyl]phenol VNPP357 (B20)3-[4-(2-benzimidazolylethyl)phenyl]benzenecarbonitrile B215-[4-(2-benzimidazolylethyl)phenyl]-2-methoxybenzenecarbonitrile B224-[4-(2-benzimidazolylethyl)phenyl]-2-cyanobenzoic acid VNPP444B(B23)1-(3′-methoxy-[1,1′-biphenyl]-4-yl)-1H-benzo[d]imidazole VNPP441C(B24)1-(3′-hydroxy-[1,1′-biphenyl]-3-yl)-1H-benzo[d]imidazole VNPP420 (B25)1-(3′-methoxy-[1,1′-biphenyl]-3-yl)-1H-benzo[d]imidazole VNPP3594′-(2-(1H-benzo[d]imidazol-1-yl)ethyl)-[1,1′-biphenyl]-3-carboxilic acidVNPP364 1-(2-(3′-(1H-tetrazol-5-yl)-[1,1′-biphenyl]-4-yl)ethyl)-1H-benzo[d]imidazoleNaphthalene Derivatives (Na Series)

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —SH, —OMe, —CN, —COON, —COOMe, —COOEt,imidazole, 1H-tetrazole and their combinations, wherein each R′ isindependently selected from alkyl or aryl.Examples of Na series

TABLE 2a Naphthalene derivatives (Na series) VNPP372/26-(3-benzimidazolylphenyl)naphthalen-2-ol (Na1) VNPP373/26-(3-benzimidazolylphenyl)-2-methoxynaphthalene (Na2) Na36-(3-benzimidazolylphenyl)naphthalene-2-carbonitrile Na46-(3-benzimidazolylphenyl)naphthalene-2-carboxylic acid Na5 methyl6-(3-benzimidazolylphenyl)naphthalene-2-carboxylate Na66-(3-benzimidazolylphenyl)-3-methoxynaphthalen-2-ol Na76-(3-benzimidazolylphenyl)-2,3-dimethoxynaphthalen Na86-(3-benzimidazolylphenyl)-3-methoxynaphthalene-2-carbonitrile Na96-(3-benzimidazolylphenyl)-3-methoxynaphthalene-2-carboxylic acid Na10methyl 6-(3-benzimidazolylphenyl)-3-methoxynaphthalene-2-carboxylateNa11 [3-(6-imidazolyl-2-naphthyl)phenyl]benzimidazole Na126-(3-benzimidazolylphenyl)-2-imidazolyl-3-methoxynaphthaleneNaphthalene Derivatives (q Series)

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —N(R′)₂—, —SH, —OMe, —CN, —COOH, —COOMe,—COOEt, imidazole, 1H-tetrazole and their combinations, wherein each R′is independently selected from alkyl or aryl.Examples of Nb Series

TABLE 2b Naphthalene derivatives (Nb series) Nb14-(5-benzimidazolyl-2-naphthyl)phenol Nb21-(5-benzimidazolyl(2-naphthyl))-4-methoxybenzene Nb34-(5-benzimidazolyl-2-naphthyl)benzenecarbonitrile Nb44-(5-benzimidazolyl-2-naphthyl)benzoic acid Nb5 methyl4-(5-benzimidazolyl-2-naphthyl)benzoate Nb64-(5-benzimidazolyl(2-naphthyl))-2-methoxyphenol Nb74-(5-benzimidazolyl(2-naphthyl))-1,2-dimethoxybenzene Nb84-(5-benzimidazolyl(2-naphthyl))-2-methoxybenzenecarbonitrile Nb94-(5-benzimidazolyl(2-naphthyl))-2-methoxybenzoic acid Nb10 methyl4-(5-benzimidazolyl(2-naphthyl))-2-methoxybenzoate Nb11[6-(4-imidazolylphenyl)naphthyl]benzimidazole Nb124-(5-benzimidazolyl(2-naphthyl))-1-imidazolyl-2-methoxybenzeneStilbene Derivatives (S Series)

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —N(R′)₂—, —SH, —OMe, —CN, —COOH, —COOMe,—COOEt, imidazole, 1H-tetrazole and their combinations, wherein each R′is independently selected from alkyl or aryl.Examples of S Series

TABLE 3 Stilbene derivatives (S series) VNPP388 (S1)4-[(1E)-2-(3-benzimidazolylphenyl)vinyl]phenol VNPP391B31-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-4-methoxybenzene (S2) S34-[(1E)-2-(3-benzimidazolylphenyl)vinyl]henzenecarbonitrile S44-[(1E)-2-(3-benzimidazolylphenyl)vinyl]benzoic acid S5 methyl4-[(1E)-2-(3-benzimidazolylphenyl)vinyl]benzoate S64-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-2-methoxyphenol S74-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-1,2-dimethoxybenzene S84-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-2-methoxybenzenecarbonitrile S94-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-2-methoxybenzoic acid S10methyl 4-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-2-methoxybenzoate S11{3-[(1E)-2-(4-imidazolylphenyl)vinyl]phenyl}benzimidazole S124-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-1-imidazolyl-2-methoxybenzeneS13 {3-[(1E)-2-(3-imidazolylphenyl)vinyl]phenyl}benzimidazole S145-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-1,2,3-trimethoxybenzeneAmide Derivatives (Amd-a Series)

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —N(R′)₂—, —SH, —OMe, —CN, —COOH, —COOMe,—COOEt, imidazole, 1H-tetrazole and their combinations, wherein each R′is independently selected from alkyl or aryl.Examples of Amd-a Series

TABLE 4a Amide derivatives (Amd-a series) Amd-a1(3-benzimidazolylphenyl)-N-(4-hydroxyphenyl)carboxamide Amd-a2(3-benzimidazolylphenyl)-N-(4-methoxyphenyl)carboxamide Amd-a3(3-benzimidazolylphenyl)-N-(4-cyanophenyl)carboxamide Amd-a44-[(3-benzimidazolylphenyl)carbonylamino]benzoic acid Amd-a5 methyl4-[(3-benzimidazolylphenyl)carbonylamino]benzoate Amd-a6(3-benzimidazolylphenyl)-N-(4-hydroxy-3-methoxyphenyl)carboxamide Amd-a7(3-benzimidazolylphenyl)-N-(3,4-dimethoxyphenyl)carboxamide Amd-a8(3-benzimidazolylphenyl)-N-(4-cyano-3-methoxyphenyl)carboxamide Amd-a94-[(3-benzimidazolylphenyl)carbonylamino]-2-methoxybenzoic acid Amd-a10methyl 4-[(3-benzimidazolylphenyl)carbonylamino]-2-methoxybenzoateAmd-a11 (3-benzimidazolylphenyl)-N-(4-imidazolylphenyl)carboxamideAmd-a12(3-benzimidazolylphenyl)-N-(4-imidazolyl-3-methoxyphenyl)carboxamideAmd-a13 (3-benzimidazolylphenyl)-N-(3-imidazolylphenyl)carboxamideAmd-a14 (3-benzimidazolylphenyl)-N-(3,4,5-trimethoxyphenyl)carboxamideAmd-a15 4-[(3-benzimidazolylphenyl)carbonylamino]-2-imidazolylbenzoicacidAmide Derivatives (Amd-b Series)

wherein is represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —N(R′)₂—, —SH, —OMe, —CN, —COOH, —COOMe,—COOEt, imidazole, 1H-tetrazole and their combinations, wherein each R′is independently selected from alkyl or aryl.Examples of Amd-b series

TABLE 4b Amide derivatives (Amd-b series) Amd-b1N-(3-benzimidazolylphenyl)(4-hydroxyphenyl)carboxamide VNPP432(Amd-N-(3-benzimidazolylphenyl)(4-methoxyphenyl)carboxamide b2) Amd-b3N-(3-benzimidazolylphenyl)(4-cyanophenyl)carboxamide Amd-b44-[N-(3-benzimidazolylphenyl)carbamoyl]benzoic acid Amd-b5 methyl4-[N-(3-benzimidazolylphenyl)carbamoyl]benzoate Amd-b6N-(3-benzimidazolylphenyl)(4-hydroxy-3-methoxyphenyl)carboxamide Amd-b7N-(3-benzimidazolylphenyl)(3,4-dimethoxyphenyl)carboxamide Amd-b8N-(3-benzimidazolylphenyl)(4-cyano-3-methoxyphenyl)carboxamide Amd-b94-[N-(3-benzimidazolylphenyl)carbamoyl]-2-methoxybenzoic acid Amd-b10methyl 4-[N-(3-benzimidazolylphenyl)carbamoyl]-2-methoxybenzoate Amd-b11N-(3-benzimidazolylphenyl)(4-imidazolylphenyl)carboxamide Amd-b12N-(3-benzimidazolylphenyl)(4-imidazolyl-3-methoxyphenyl)carboxamideAmd-b13 N-(3-benzimidazolylphenyl)(3-imidazolylphenyl)carboxamideAmd-b14 N-(3-benzimidazolylphenyl)(3,4,5-trimethoxyphenyl)carboxamideAmd-b15 4-[N-(3-benzimidazolylphenyl)carbamoyl]-2-imidazolylbenzoic acidAmine Derivatives (Am-a Series)

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NM′, —SH, —OMe, —CN, —COOH, —COOMe, —COOEt,imidazole, 1H-tetrazole and their combinations, wherein each R′ isindependently selected from alkyl or aryl.Examples of Am-a series

TABLE 5a VNPP428(Am- 4-{[(3-benzimidazolylphenyl)amino]methyl}phenol a1)VNPP429(Am- (3-benzimidazolylphenyl)[(4-methoxyphenyl)methyl]amine a2)Am-a3 4-{[(3-benzimidazolylphenyl)amino]methyl}benzenecarbonitrile Am-a44-{[(3-benzimidazolylphenyl)amino]methyl}benzoic acid Am-a5 methyl4-{[(3-benzimidazolylphenyl)amino]methyl}benzoate Am-a64-{[(3-benzimidazolylphenyl)amino]methyl}-2-methoxyphenol Am-a7(3-benzimidazolylphenyl)[(3,4-dimethoxyphenyl)methyl]amine Am-a84-{[(3-benzimidazolylphenyl)amino]methyl}-2- methoxybenzenecarbonitrileAm-a9 4-{[(3-benzimidazolylphenyl)amino]methyl}-2-methoxybenzoic acidAm-a10 methyl4-{[(3-benzimidazolylphenyl)amino]methyl}-2-methoxybenzoate Am-a11(3-benzimidazolylphenyl)[(4-imidazolylphenyl)methyl]amine Am-a12(3-benzimidazolylphenyl)[(4-imidazolyl-3-methoxyphenyl)methyl]amineAm-a13 (3-benzimidazolylphenyl)[(3-imidazolylphenyl)methyl]amine Am-a14(3-benzimidazolylphenyl)[(3,4,5-trimethoxyphenyl)methyl]amine Am-a154-{[(3-benzimidazolylphenyl)amino]methyl}-2-imidazolylbenzoic acidAmine Derivatives (Am-b Series)

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —N(R′)₂—, —SH, —OMe, —CN, —COOH, —COOMe,—COOEt, imidazole, 1H-tetrazole and their combinations, wherein each R′is independently selected from alkyl or aryl.Examples of Am-a Series

TABLE 5b Amine derivatives (Am-b series) Am-b14-{[(3-benzimidazolylphenyl)methyl]amino}phenol Am-b2[(3-benzimidazolylphenyl)methyl](4-methoxyphenyl)amine Am-b34-{[(3-benzimidazolylphenyl)methyl]amino}benzenecarbonitrile Am-b44-{[(3-benzimidazolylphenyl)methyl]amino}benzoic acid Am-b5 methyl4-{[(3-benzimidazolylphenyl)methyl]amino}benzoate Am-b64-{[(3-benzimidazolylphenyl)methyl]amino}-2-methoxyphenol Am-b7[(3-benzimidazolylphenyl)methyl](3,4-dimethoxyphenyl)amine Am-b84-{[(3-benzimidazolylphenyl)methyl]amino}-2-methoxybenzenecarbonitrileAm-b9 4-{[(3-benzimidazolylphenyl)methyl]amino}-2-methoxybenzoic acidAm-b10 methyl4-{[(3-benzimidazolylphenyl)methyl]amino}-2-methoxybenzoate Am-b11[(3-benzimidazolylphenyl)methyl](4-imidazolylphenyl)amine Am-b12[(3-benzimidazolylphenyl)methyl](4-imidazolyl-3-methoxyphenyl)amineAm-b13 [(3-benzimidazolylphenyl)methyl](3-imidazolylphenyl)amine Am-b14[(3-benzimidazolylphenyl)methyl](3,4,5-trimethoxyphenyl)amine Am-b154-{[(3-benzimidazolylphenyl)methyl]amino}-2-imidazolylbenzoic acidSulphonamide Series—SulAmd Series

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —N(R′)₂—, —SH, —OMe, —CN, —COOH, —COOMe,—COOEt, imidazole, 1H-tetrazole and their combinations, wherein each R′is independently selected from alkyl or aryl.Examples of SulAmd Series

TABLE 6 Sulphonamide series - SulAmd series SulAmd1N-(3-benzimidazolylphenyl)[(4-hydroxyphenyl)sulphonyl]amine VNPP431N-(3-benzimidazolylphenyl)[(4-methoxyphenyl) sulphonyl]amine (SulAmd2)SulAmd3 N-(3-benzimidazolylphenyl)[(4-cyanophenyl) sulphonyl]amineSulAmd4 4-[N-(3-benzimidazolylphenyl)Sulphonamyl]benzoic acid SulAmd5methyl 4-[N-(3-benzimidazolylphenyl)Sulphonamyl]benzoate SulAmd6N-(3-benzimidazolylphenyl)[(4-hydroxy-3-methoxyphenyl) sulphonyl]amineSulAmd7 N-(3-benzimidazolylphenyl)[(3,4-dimethoxyphenyl) sulphonyl]amineSulAmd8 N-(3-benzimidazolylphenyl)[(4-cyano-3-methoxyphenyl)sulphonyl]amine SulAmd94-[N-(3-benzimidazolylphenyl)Sulphonamyl]-2-methoxybenzoic acid SulAmd10methyl 4-[N-(3-benzimidazolylphenyl)Sulphonamyl]-2-methoxybenzoateSulAmd11 N-(3-benzimidazolylphenyl)[(4-imidazolylphenyl) sulphonyl]amineSulAmd12 N-(3-benzimidazolylphenyl)[(4-imidazolyl-3-methoxyphenyl)sulphonyl]amine SulAmd13 N-(3-benzimidazolylphenyl)[(3-imidazolylphenyl)sulphonyl]amine SulAmd14N-(3-benzimidazolylphenyl)[(3,4,5-trimethoxyphenyl) sulphonyl]amineSulAmd15 4-[N-(3-benzimidazolylphenyl)Sulphonamyl]-2-imidazolylbenzoicacidTetrahydroisoquinoline Derivatives (Q Series)

wherein R represents mono or di substitutions selected from the groupconsisting of —OH, —NH₂, —NHR′, —SH, —OMe, —CN, —COOH, —COOMe, —COOEt,imidazole, 1H-tetrazole and their combinations, wherein each R′ isindependently selected from alkyl or aryl. The squiggly line representsan α or β-methyl group that may be either projecting in front of theplane of the paper or projecting behind the plane of the paper.Examples of Q series

TABLE 7 Tetrahydroisoquinoline derivatives (Q series) Q12-[(3-benzimidazolylphenyl)methyl]-3-methyl-1,2,3,4-tetrahydroisoquinolin-6-ol(Q1) Q2 2-[(3-benzimidazolylphenyl)methyl]-6-methoxy-3-methyl-1,2,3,4-tetrahydroisoquinoline (Q2) Q32-[(3-benzimidazolylphenyl)methyl]-3-methyl-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile (Q3) Q42-[(3-benzimidazolylphenyl)methyl]-3-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (Q4) Q5 methyl2-[(3-benzimidazolylphenyl)methyl]-3-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylate (Q5) Q62-[(3-benzimidazolylphenyl)methyl]-7-methoxy-3-methyl-1,2,3,4-tetrahydroisoquinolin-6-ol (Q6) Q72-[(3-benzimidazolylphenyl)methyl]-6,7-dimethoxy-3-methyl-1,2,3,4-tetrahydroisoquinoline (Q7) Q82-[(3-benzimidazolylphenyl)methyl]-7-methoxy-3-methyl-1,2,3,4-tetrahydroisoquinoline-6-carbonitrile (Q8) Q92-[(3-benzimidazolylphenyl)methyl]-7-methoxy-3-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylic acid (Q9) Q10 methyl2-[(3-benzimidazolylphenyl)methyl]-7-methoxy-3-methyl-1,2,3,4-tetrahydroisoquinoline-6-carboxylate (Q10) Q11{3-[(6-imidazolyl-3-methyl-2-1,2,3,4-tetrahydroisoquinolyl)methyl]phenyl}benzimidazole (Q11) Q122-[(3-benzimidazolylphenyl)methyl]-6-imidazolyl-7-methoxy-3-methyl-1,2,3,4-tetrahydroisoquinoline (Q12) Q13 {3-[(7-imidazolyl-3-methyl-2-1,2,3,4-tetrahydroisoquinolyl)methyl]phenyl}benzimidazole (Q13) Q142-[(3-benzimidazolylphenyl)methyl]-5,6,7-trimethoxy-3-methyl-1,2,3,4-tetrahydroisoquinoline (Q14)Androstene Derivatives

The two squiggly lines attached directly to the compound A represent asubstituent group that may be either projecting in front of the plane ofthe paper (example (A1) or projecting behind the plane of the paper(example Example A3).

The two squiggly lines on the right side of each partial structure beloware an abbreviation for the rest of the andorostene derivative molecule.See formula A.

Examples of Androstene Derivatives (A)

TABLE 8 Androstene derivatives VNPP₄₃₃-3β3β-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(2d)(A1) VNPP4333β-(1H-benzimidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(A2) VNPP433-3α3α-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene (A3)A43α-(1H-benzimidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(A4) VNPP3973β-(1H-imidazole-1-thiocarboxylate)-17-(1H-benzimidazol-1-yl)androsta-(A5) 5,16-diene A63β-(1H-benzimidazole-1-carboxylate)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A73β-(1H-benzimidazole-1-thiocarboxylate)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A83β-Thiol-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A93β-(1H-imidazole-1-carbothioate)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A103β-(1H-imidazole-1-carbodithioate)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A113β-(pyridin-2-yloxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A123β-(pyridin-3-yloxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A133β-(pyridin-4-yloxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A143β-(pyridin-2-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneVNPP415C3β-(pyridin-3-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(A15) VNPP4143β-(pyridin-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(A16) A173β-(thiazol-5-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneA183β-(thiazol-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneA193β-(thiazol-2-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneA20 3β-(thiazol-5-yloxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneA21 3β-(thiazol-4-yloxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneA22 3β-(thiazol-2-yloxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneA23 3β-(picolinamide)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A243β-(nicotinamide)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A253β-(isonicotinamide)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A263β-(nitrile)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A273β-(carboxylic acid)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A283β-(methylcarboxylate)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A293β-(ethylcarboxylate)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene A303β-(1H-tetrazol-5-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneVNPP412 3β-(Methoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene (A31)A323β-(Imidazol-1-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-dieneEstrogen Derivatives

The two squiggly lines attached directly to the compound A represent asubstituent group that may be either projecting in front of the plane ofthe paper (example (A1) or projecting behind the plane of the paper(example Example A3).

The two squiggly lines on the right side of each partial structure beloware an abbreviation for the rest of the andorostene derivative molecule.See formula A.

Examples of Estrogen Derivatives

TABLE 9 Estrogen derivatives E13-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E23-(1H-benzimidazol-1-yl)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE3 3-(nitrile)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E43-(1H-tetrazol-5-yl)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienVNPP341 3-(carboxylicacid)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien (E5) VNPP3343-(methylcarboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien(E6) E73-(ethylcarboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E83-(1H-imidazol-1-carboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E93-(1H-imidazol-1-carbothioate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E103-(1H-benzimidazol-1-carboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E113-(1H-benzimidazol-1-carbothioate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E123-(pyridine-2-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE133-(pyridine-3-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE143-(pyridine-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE153-(thiazol-5-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE163-(thiazol-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE173-(thiazol-2-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE18 3-(thiol)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien VNPP4052-(methoxy)-3-(hydroxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien(E19) E202-(methylthio)-3-(hydroxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienVNPP338 3-(hydroxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien(E21) E22 2-(methoxy)-3-(carboxylicacid)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)- trien E232-(methoxy)-3-(methylcarboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E242-(methoxy)-3-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E252-(methoxy)-3-(1H-benzimidazol-1-yl)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E262-(methoxy)-3-(nitrile)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE272-(methoxy)-3-(1H-tetrazol-5-yl)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E282-(methoxy)-3-(ethylcarboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E292-(methoxy)-3-(1H-imidazol-1-carboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E302-(methoxy)-3-(1H-imidazol-1-carbothioate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E312-(methoxy)-3-(1H-benzimidazol-1-carboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E322-(methoxy)-3-(1H-benzimidazol-1-carbothioate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E332-(methoxy)-3-(pyridine-2-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E342-(methoxy)-3-(pyridine-3-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E352-(methoxy)-3-(pyridine-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E362-(methoxy)-3-(thiazol-5-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E372-(methoxy)-3-(thiazol-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E382-(methoxy)-3-(thiazol-2-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien E392-(methoxy)-3-(thiol)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trienE403-(imidazol-1-ylmethoxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien

EXPERIMENTAL SECTION

General:

All chemicals and reagents were commercially obtained from Aldrich orAcros chemicals except 6-hydroxynaphthyl-2-boronicacidacid (Alfa Aesar)and are used directly without purification. Reaction progress wasdetermined by TLC analysis on Silica gel plates (Merck F254). FlashColumn chromatography (FCC) was performed using silica gel (230-400mesh, 60 Å). Melting points (mp) were determined with a Stuart SMP10melting point apparatus and are uncorrected. IR spectra were recordedneat on a Perkin Elmer spectrum65 FT IR spectrometer and are reported asλmax (cm⁻¹). ¹H NMR spectra were recorded on a Bruker DMX 500 MHz orBruker Ascend 400 MHz spectrometer using CDCl₃ or DMSO-d₆ as solvent.Chemical shifts are given in parts per million (ppm), and TMS was usedas an internal standard. ¹³C NMR spectra were recorded Bruker DMX 500MHz or Bruker 400 MHz spectrometer operating at 125 or 100 MHzrespectively.

Chemistry:

The syntheses of all compounds in this study were accomplished byfollowing methods and conditions as depicted in Scheme 1-8. The keyintermediate N-alkylated benzimidazoles (VNPP319C and VNPP346) for thesynthesis biphenyl derivatives were prepared by reacting benzimidazolewith appropriate arylalkylbromide, in presence KOH in dry DMSO withexcellent yield (Method A1, Scheme 1).²⁶ Where, N-arylated bezimidazole(VNPP371/1, VNPP423 and VNPP442) were obtained in moderate yield byapplying reported method of ‘ligated catalysis’ for the Ullmanncondenstation (Method A2, Scheme 1).²⁷ In which, benzimidazole isreacted with substituted 3-iodobenzenes in presence of mild alkali(cesium carbonate), catalyst (copper iodide) and a ligate(1,10-phenantroline) in DMF at 110° C. for 40 h. These alkyl/arylatedbenzimidazoles were reacted with corresponding substituted boronic acidsvia Suzuki coupling to form desired biaryl (VNPP347B(B1), VNPP358(B2),VNPP356(B3), VNPP360(B7), VNPP321(B18), VNPP355(B19), VBPP357(B20),VNPP444B(B23), VNPP441C(B24), VNPP420(B25), VNPP359,VNPP372/2(Na1)—VNPP373/2(Na2)) compounds (Method B, Scheme 1).²³ Further1,3-diploar cycloaddition of sodium azide with nitrile biphenyls(VNPP358 & 357) in presence of ammonium chloride in polar high-boilingsolvent such as DMF afforded excellent yield of respective tetrazolecompounds (VNPP361(B11) and VNPP364; Method C, Scheme 1).²⁵

Synthesis of stilbene derivative with mainly (E)-configuration(VNPP388(S1) and VNPP391B3(S2)) are achieved with moderate yield, inessence, as described in recent report (Scheme 2).²⁸ Thus, reaction ofaldehyde with methyltriphenylphosphonium bromide in the presence oft-BuOK in a Wittig method provided the intermediate styrene (VNPP383).Its Mizoroki-Heck reaction with bromoaryl benzimidazole (VNPP371/1) intriethanolamine as a base/ligand/solvent in presence of catalytic amountof Pd(OAc)₂ at 100° C. for 24 h provided stilbene (VNPP388(S1)) in goodyield. Similarly compound VNPP390(S2) is synthesized from commerciallyavailable p-methoxyphenylstyrene.

Diaryl compounds with alkylamines (VNPP428(Am-a1), VNPP429(Am-a2)),amide (VNPP432(Amd-b2)) and sulfonamide (VNPP431(SulAmd-1)) linkercompounds were synthesized using arylated BzIm intermediate1-(3-aminophenyl-1H-benzo[d]imidazole (VNPP423) compound (Scheme 3). Theintermediate VNPP423 was synthesized using 3-iodoaniline by following(Method-A2) procedure for intermediate VNPP371.²⁷ Alkylamine(VNPP428(Am-a1)-VNPP429(Am-a2))compounds were obtained via imines (notisolated) by refluxing VNPP423 with corresponding aldehyde in ethanol inpresence of molecular sieves and further reducing imines with sodiumborohyderide in methanol at ice cold temperature (34% and 50%respectively).²⁵ The amide derivative VNPP432(Amd-b2) is obtained by thereaction of 4-methoxybenzoyl chloride on VNPP423in presence TEA in ethylacetate at room temperature (49%). Where sulfonamide derivativeVNPP431(SulAmd-1) is achieved by refluxing VNPP423 with4-methoxybenzenesulfonyl chloride in pyridine (48%).²⁵

For the synthesis of C3 imidazole derivatives of VN/124-1, initiallymesyl derivative of VN/124-1 (VNPT88) prepared (Scheme 4) which onrefluxing with imidazole in toluene under anhydrous condition resultedin three imidazole substituted compounds (VNPP433-6 (35%),VNPP433-3α(A3) (3%) and VNPP433-3β(2d)(A1) (11%)). These threepositional isomers were separated by preparative HPLC method. Thethiocarbamate derivative VNPP397(A5) is accomplished by slightlymodifying our reported synthetic method of (Scheme 5) imidazolecarbamatederivative of VN/124-1.¹⁷ Thus VN/124-1 is refluxed with1,1′-thiocarbonyldiimidazole in acetonitrile and MDC solvent mixture.The C3 ether derivatives (VNPP415C(A15), VNPP414(A16) and VNPP412(A31))of VN/124-1 were prepared by following williamson's etherfication(Scheme 6) method of treating alcohol (VN/124-1) with arylalkyl/alkylhalide in presence of sodium hydride as base in DMF.

The synthesis of 17-bezoazole of estrone-3-carboxlate (VNPP341(E5)) andestrone (VNPP338(E21)) derivatives is outlined in scheme 7 and 8,respectively. For the synthesis of estrone-3-carboxylate derivative(VNPP341(E5), initially 3-enol carbon of estrone is activated byconverting it in to triflate (VNPP308) by reacting with triflicanhydride in presence of organic base (TEA).²⁹ Then triflate is replacedwith methyl carboxylate through palladium-catalyzed carbonylation usingPd(OAc)₂ as catalyst, 1,c-bis(diphenylphospheno)propane as the phosphineligand, in presence of gaseous carbon monoxide, methanol in DMF(VNPP309A).³⁰ The synthesis of title compound 17-1H-bezimidazolylderivative (VNPP341(E5) of this estrone-3-carboxylate, is accomplishedby following our routine method for the synthesis of VN/124-1 as shownin Scheme 7.¹⁷ Which follows three intermediate steps viz formation of16-formyl-17-bromo derivative (VNPP315B) through vilsmeier-hackreaction, condensation of bezimidazole ring to C-17 position (VNPP330),then Pd catalyzed 16-deformylation (VNPP334(E6)) and finally basichydrolysis of methyl ester group to obtain target compound VNPP341(E5).

The synthesis of estrone-3-hydroxy-17-1H-bezoazole (VNPP334(E21))derivative initiated with 3-acetylation of commercially availableestrone (Scheme 8) with acetic anhydride in pyridine with 98% ofisolated yield (VNPP310). Then it is subjected to vilsmeier-hackreaction to obtain 16-formy-17-bromo derivative (VNPP311) by treatingwith phosphorus tribromide in DMF and chloroform with low yield. Anattempt to condensation of benzimidazole to 17^(th) position of VNPP311by following our routine K₂CO₃ in DMF method resulted into mixture offour components (partial 3-deacetylation with or without benzimidazolesubstitution, acetylated benzimidazole product and substrate).Therefore, first the 3-acetyl group of VNPP311 was hydrolyzed bytreating with10% ethanolic-KOH to obtain VNPP312, then benzimidazolegroup condensed using K₂CO₃ in DMF (VNPP314) and finally16-deformylation by refluxing with 10% Pd/C in benzonitrile to obtainfinal compound (VNPP338(E21)) with very low isolated yield in the finalstep (6%).

General Methods for Synthesis:Method A1: N-alkylation of Benzimidazole (VNPP319C and VNPP346).

DMSO (10 mL, dried over molecular sieves) was added to KOH (0.95 g,16.96 mmol, crushed pallets) and the mixture was stirred for 5 min.Benzimidazole (0.5 g, 4.23 mmol) was then added and stirred at roomtemperature for 2 h prior to the addition of corresponding4-bomophenylakyl bromide (8.46 mmol). Reaction mixture stirred for 24 h,poured on to ice-water mixture, filtered, dried and purified by FCC

Method A2: Arylation of Benzimidazole (Compound VNPP442, VNPP371/1 andVNPP423)

A mixture of benzimidazole (1 equiv), corresponding substitutediodobenzene (1.2 equiv), copper iodide (0.01 equiv), 1,10-phenanthroline(0.2 equiv) and cesium carbonate (2 equiv) in DMF was heated to 110° C.for 40 h. The reaction mixture was cooled to room temperature and thesolvent was evaporated under vacuum. Residue stirred with water andEtOAc, filtered, organic layer separated, dried (Na₂SO₄) and the solventremoved under vacuum. Crude product was purified by FCC using gradientsolvent [petroleum ether/EtOAc (8:2) then (1:1) and traces of TEA].

Method B: Suzuki Coupling.

The corresponding brominated aromatic compound (1 equiv) was dissolvedin toluene (7 mL/mmol), and to this an aqueous 2.0 M Na₂CO₃ solution(3.2 mL/mmol), corresponding boronic acid (1.5-2 equiv), ethanol (3.2mL/mmol of boronic acid) and tetrabutylammonium bromide (1 equiv) wereadded. The mixture was deoxygenated under vacuum and flushed with argon.After this cycle was repeated five times, Pd(OAc)₂ (5 mol %) was addedand the resulting suspension was refluxed for 2-6 h. Reaction mixturecooled, EtOAc (10 mL) and water (10 mL) were added and the organic phasewas separated. The water phase was extracted with EtOAc (2×10 mL). Thecombined organic phases were washed with brine, dried over Na₂SO₄,filtered over a short plug of Celite, and evaporated under vacuum. Thecompounds were purified by FCC on 5 inch silica gel column usingpetroleum ether/EtOAc (1:1) to remove nonpolar contaminations then with2% methanol in DCM. Most of biphenyls were synthesized, isolated andpurified by this method unless otherwise mentioned.

Method C: Conversion of Nitrile to Tetrazole.

A mixture of corresponding nitrile compound (1 equiv), sodium azide(12.5 equiv) and ammonium chloride (12.5 equiv) in DMF was heated at120° C. for 20 h. The reaction mixture was poured into ice cold water(30 mL) and acidified with dilute HCl solution. The white precipitateobtained on acidification was filtered; dried and aqueous phase wasextracted with EtOAc, dried with Na₂SO₄ and evaporated. Crude solid andcrude product obtained by EtOAc extraction were combined, purified byFCC over a short column [10% methanol in DCM] to give pure tetrazolederivative.

Method D: Amine Derivatives (VNPP428(am-A1), VNPP429(am-a2))

A mixture of amine VNPP423 (1 equiv), corresponding aldehyde (1 equiv),and molecular sieves in ethanol refluxed overnight. Then solventevaporated, reconstituted with methanol and NaBH₄ (2 equiv) added at icecold temperature and stirred for 1 hr. Reaction mixture filtered andthen purified by FCC [petroleum ether/EtOAc/TEA (3:2:0.01)].

Experimental 1-(4-bromophenethyl)-1H-benzo[d]imidazole (VNPP319C)

Method A1: using KOH (0.95 g, 16.96 mmol), DMSO (10 ml) benzimidazole(0.5 g, 4.23 mmol) and 4-bomophenylethyl bromide (2.23 g, 8.46 mmol).Crude product was purified by FCC using short column [2% methanol inDCM] to give VNPP319C (1.05 g, 82.7%): mp 124-126° C.; IR (Neat) 1610,1500, 1486, 1456, 1242, 1168, 1069, 1008, 805, 747 cm⁻¹; ¹H NMR (500MHz, CDCl₃) δ 3.10 (t, J=6.94 Hz, 2 H, CH₂), 4.38 (t, J=6.87 Hz, 2 H,═N—CH₂), 6.86 (d, J=8.09 Hz, 2 H, Ar-2² and 6² Hs), 7.27-7.34 (m, 2 H,Ar-5¹ and 6¹ Hs), 7.35-7.41 (comp, 3 H, Ar-7¹, 3 ² and 5² Hs), 7.58 (s,1 H, Ar-2¹ H), 7.77-7.85 (dd, J=6.65, 1.75 Hz, 1 H, Ar-4¹ H); ¹³C NMR(101 MHz, CDCl₃) δ 35.6, 46.4, 109.4, 120.6, 121.0, 122.2, 123.0, 130.3(2×C), 131.9 (2×C), 133.4, 136.4, 142.9, 143.9

1-(4-bromobenzyl)-1H-benzo[d]imidazole (VNPP346)

Method A1: using KOH (0.95 g, 17 mmol), DMSO (10 ml), benzimidazole (0.5g, 4.23 mmol) and 4-bomobenzyl bromide (2.11 g, 8.46 mmol). Crudeproduct was purified by FCC using short column [6% EtOAc in petroleumether and then 20%] to give VNPP346 (1.0 g, 82.6%): mp 87-91° C.; IR(Neat) 1591, 1617, 1488, 1468, 1364, 1285, 1260, 1205, 1172, 1099, 739cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ 5.32 (s, 2 H, CH₂), 7.05 (d, 2 H, J=8.24Hz, Ar-2² and 6² Hs), 7.24-7.27 (comp, 3 H, Ar-5¹, 6¹ and 7¹-Hs), 7.47(d, 2 H, J=8.39 Hz, Ar-3², 5² Hs), 7.83 (d, 1 H, J=7.48 Hz, Ar-4¹ H),7.95 (s, 1 H, Ar-2¹ H); ¹³C NMR (101 MHz, CDCl₃) δ 110.2, 120.8, 122.5,123.1, 123.5, 124.0, 127.0, 131.0, 131.3, 133.3, 137.5, 141.9, 144.0

1-(4-bromophenyl)-1H-benzo[d]imidazole (VNPP442)

Method A2: using 3-bromoiodobenzene. Yield 14%: mp 100-1° C.; ¹H NMR(400 MHz, CDCl₃) δ 7.33-7.37 (m, 2 H, Ar-5¹ and 6¹ Hs), 7.39-7.44 (d, 2H, Ar-2², 6² Hs), 7.48-7.56 (m, 1 H, Ar-7¹), 7.69-7.74 (d, 2 H, Ar-3²,5²Hs), 7.86-7.91 (m, 1 H, Ar-4¹), 8.08 (s, 1 H, Ar-2¹); ¹³C NMR (500MHz, CDCl₃) δ 144.11, 141.98, 135.41, 133.48, 133.27, 125.55, 123.97,123.05, 121.66, 120.81, 110.22

1-(3-bromophenyl)-1H-benzo[d]imidazole (VNPP371/1)

Method A2 using 3-bromoiodobenzene. Yield 60.8%: mp 77-79° C.; ¹H NMR(500 MHz, DMSO-d₆) δ 7.34 (quin, J=7.40 Hz×4, 2 H, Ar-5¹ and 6¹Hs),7.56-7.62 (m, 1 H, 5² H), 7.64 (d, J=7.48 Hz, 1 H Ar-6² H), 7.71 (d,J=8.09 Hz, 1 H, Ar-7¹ H), 7.74 (d, J=7.93 Hz, 1 H, Ar-4² H), 7.79 (d,J=7.48 Hz, 1 H Ar-4¹ H), 7.96 (s, 1 H, Ar-2¹ H), 8.60 (s, 1 H, Ar-2² H);¹³C NMR (500 MHz, CDCl₃) δ 144.2, 142.1, 137.7, 133.5, 131.5, 131.3,127.2, 124.2, 123.7, 123.3, 122.7, 120.9, 110.4

1-(3-aminophenyl)-1H-benzo[d]imidazole (VNPP423)

Method A2: using 3-iodoniline. Yield 78%: mp 137-38° C.; ¹H NMR (400MHz, CDCl₃) δ 3.94 (br. s., 2 H, NH₂), 6.72-6.77 (m, 1 H, Ar-5² H), 6.78(s, 1 H, Ar-2² H), 6.87 (d, J=7.34 Hz, 1 H, Ar-4² H), 7.27-7.40 (m, 3 H,Ar-6², 5¹ and 6¹ Hs), 7.54-7.63 (m, 1 H, Ar-4¹), 7.81-7.92 (m, 1 H,Ar-7¹ H), 8.09 (s, 1 H, Ar-2¹ H); ¹³C NMR (500 MHz, CDCl₃) δ 148.0,142.3, 137.3, 133.6, 130.8, 123.5, 122.6, 120.5, 114.4, 113.7, 110.7,110.1

3-[4-(benzimidazolylmethyl)phenyl]-1-methoxybenzene (VNPP347B (B1))

Method B: using VNPP346 (0.3 g, 1.05 mmol) and 3-methoxyphenylboronicacid (0.24 g, 1.57 mmol). Yield (0.31 g, 94%): mp 132-133° C.; IR(Neat2960, 1607, 1584, 1567, 1482, 1437, 1296, 1208, 1174, 1058, 1014,839, 777 cm⁻¹; ¹H NMR (500 MHz, CDCl3) δ 3.87 (s, 3 H, CH₃), 5.43 (s, 2H, CH₂), 6.92 (dd, J=8.24, 1.98 Hz, 1 H, Ar-4³ H), 7.09 (s, 1 H, Ar-2³H), 7.15 (d, J=7.78 Hz, 1 H, Ar-6³ H), 7.25-7.28 (m, 2 H, Ar-5¹ and 6¹Hs), 7.28-7.32 (m, 2 H, Ar-2² and 6² Hs), 7.32-7.39 (m, 2 H, Ar-5³ and7¹ Hs), 7.57 (d, J=8.09 Hz, 2 H, Ar-3² and 5² H), 7.86 (d, J=7.78 Hz, 1H, Ar-4¹ H), 8.01 (s, 1 H, Ar-2¹-H); ¹³C NMR (500 MHz, CDCl₃) δ 160.2,144.1, 143.4, 142.0, 141.3 134.7, 134.1, 130.0, 127.9 (2×C), 127.7(2×C), 123.3, 122.5, 120.6, 119.7, 113.1, 113.0, 110.3, 55.5, 48.7

3-[4-(benzimidazolylmethyl)phenyl]benzenecarbonitrile (VNPP358 (B2))

Method B: using VNPP346 (0.19 g, 0.66 mmol) and 3-nitrilephenylboronicacid (0.15 g, 1 mmol). Yield (0.18 g, 86.7%): mp 133-135° C.; IR (Neat)2230, 1491, 1458, 1368, 1286, 1260, 1183, 796 cm⁻¹; ¹H NMR (500 MHz,D_(MS)O-d₆) δ 5.57 (s, 2 H, CH₂), 7.17-7.25 (m, 2 H, Ar-5¹ and 6¹ Hs),7.43 (m, J=8.24 Hz, 2 H, Ar-2² and 6² Hs), 7.54-7.59 (m, 1 H, Ar-7¹ H),7.65 (m, 2 H, Ar-4¹ and 5³ H), 7.72 (m, 2 H, Ar-3² and 5² Hs), 7.81 (d,J=7.78 Hz, 1 H, Ar-6³ H), 7.99 (d, J=8.70 Hz, 1 H, Ar-64H), 8.12 (s, 1H, Ar-2² H), 8.46 (s, 1 H, Ar-2³ H); ¹³C NMR (500 MHz, CDCl₃) δ 144.1,143.5, 141.7, 139.0, 136.0, 134.0, 131.6, 131.1, 130.8, 129.9, 128.0,127.9, 123.5, 122.6, 120.7, 118.9, 113.2, 110.2, 48.6

3-[4-(benzimidazolylmethyl)phenyl]phenol (VNPP356 (B3))

Method B: using VPNN346 (0.20 g, 0.7 mmol) and 3-hydroxyphenylboronicacid (0.15 g, 1.08 mmol). Yield (0.14 g, 65.7%): mp 231-232° C.; IR(Neat) 3103, 1582, 1567, 1504, 1476, 1364, 1305, 1219, 1196, 830,777cm⁻¹; ¹H NMR (500 MHz, DMSO-d₆) δ 5.54 (br. s., 2H, CH₂), 6.77 (d,J=7.32 Hz, 1 H, Ar-4³ H), 7.00 (br. s., 1 H, Ar-2³ H), 7.03 (d, J=7.48Hz, 1 H, Ar-6³ H), 7.22 (m, J=7.02 Hz, 3 H, Ar-5¹, 6¹ and 5³Hs), 7.38(d, J=7.48 Hz, 2 H, Ar-2² and 6² Hs), 7.57 (d, J=7.02 Hz, 3 H, Ar-3², 5²and 7¹ Hs), 7.69 (d, J=6.87 Hz, 1 H, Ar-4¹ H), 8.46 (s, 1 H, Ar-2¹ H),9.56 (s, 1 H, —OH); ¹³C NMR (500 MHz, DMSO-d₆) δ 158.4, 144.9, 144.2,141.7, 140.4, 136.7, 134.3, 130.5, 128.5 (2×C), 127.5 (2×C), 123.1,122.2, 120.1, 118.0, 115.2, 114.1, 111.3, 47.9

3-[4-(benzimidazolylmethyl)phenyl]benzoic acid (VNPP360 (B7))

By slight modification of Method B: using VNPP346 (0.19 g, 0.66 mmol)and 3-carboxyphenylboronic acid (0.17 g, 1 mmol) and 2 M aq Na₂CO₃(2×2.13 mL). After completion of reaction, reaction mixture neutralizedwith dil. HCl, extracted with EtOAc, filtered and concentrated. Onpurification by FCC [petroleum ether/EtOAc (1:1) and then with 4%ethanol in DCM with traces of CH₃COOH] gave VNPP360 (0.03 g, 14%): mp240-242° C.; IR (Neat) 3337, 1615, 1499, 1461, 1266, 1207, 806, 735cm⁻¹¹H NMR (500 MHz, DMSO-d₆) δ 5.57 (s, 2 H, CH₂), 7.21 (quind,J=7.17×4, 1.37 Hz, 2 H, Ar-5¹ and 6¹ Hs), 7.42 (d, J=8.24 Hz, 2 H, Ar-2²and 6² Hs), 7.52-7.58 (m, 2 H, Ar-4¹ and 7¹Hs), 7.67 (m, 3 H, Ar-5³, 3²and 5² Hs), 7.84 (d, J=8.09 Hz, 1 H, Ar-6³ H), 7.91 (d, J=7.78 Hz, 1H,Ar-4³ H), 8.14 (s, 1 H, Ar-2¹ H), 8.45 (s, 1 H, Ar-2³ H); ¹³C NMR (500MHz, DMSO-d₆) δ 168.2, 144.2, 140.4, 139.4, 137.2, 134.3, 133.0, 131.2,129.8, 129.0, 128.8, 128.7 (2×C), 128.0, 127.7 (2×C), 122.4, 122.1,120.1, 111.3, 47.9

5-{3-[4-(benzimidazolylmethyl)phenyl]phenyl}-1H-1,2,3,4-tetraazole(VNPP361 (B11))

Method C: using 0.85 g (0.274 mmol) of VNPP358. Yield (0.075 g, 77.4%):mp 231-233° C.; IR (Neat) 3337, 1615, 1499, 1461, 1266, 1207, 806, 735cm¹¹H NMR (500 MHz, DMSO-d₆) δ 5.59 (s, 2 H, CH₂), 7.22 (quin, J=7.13Hz, 2 H, Ar-5¹ and 6¹ Hs), 7.45 (d, J=7.78 Hz, 2 H, Ar-2² and 2⁶ Hs),7.59 (d, J=6.71 Hz, 2 H, Ar-3² and 5² Hs), ), 7.68 (d, J=7.32 Hz, 1 H,Ar-6³ H), 7.70-7.79 (m, 3 H, Ar-7¹, 5³, and 4¹), 8.07 (d, J=7.48 Hz, 1H, Ar-4³ H), 8.40 (br. s., 1 H, Ar-2¹ H), 8.49 (br. s., 1 H, Ar-2³ H);¹³C NMR (500 MHz, DMSO-d₆) δ 157.7, 144.9, 144.2, 140.8, 139.6, 137.2,134.3, 130.3, 128.7 (2×C), 128.3, 127.7 (2×C), 126.3, 125.4, 124.6,123.1, 122.2, 120.1, 111.4, 47.9

3-[4-(2-benzimidazolylethyl)phenyl]-1-methoxybenzene (VNPP321(B18))

Method B: using VNPP319C (0.2 g, 0.66 mol) and 3-methoxyphenylboronicacid (0.16 g, 1.05 mmol). On purification by FCC over short column[petroleum ether/EtOAc (1:1)] gave VNPP321 as viscous liquid (0.16 g,73%): IR (Neat) 2936, 1599, 1480, 1457, 1288, 1214, 1168, 1051, 778, 741cm⁻¹; ¹H NMR (500 MHz, CDCl₃) 67 3.16 (t, J=7.02 Hz, 2 H, CH₂), 3.85 (s,3 H, CH₃), 4.41 (t, J=7.02 Hz, 2H, ═N—CH₂), 6.88 (dd, J=8.24, 1.83 Hz, 1H, Ar-4³-H), 7.05-7.10 (m, 3 H, Ar-6³, 5¹ and 6¹ Hs), 7.13 (d, J=7.63Hz, 1 H, Ar-7¹-H), 7.28-7.36 (m, 3 H, Ar-2², 6² and 2³ Hs), 7.37-7.42(m, 1 H, Ar-5³-H), 7.48 (d, J=8.09 Hz, 2 H, Ar-3² and 5²Hs). 7.66 (s, 1H, Ar-2¹-H), 7.82 (dd, J=6.18, 2.52 Hz, 1 H, Ar-4¹ H);¹³C NMR (500 MHz,CDCl₃) δ 160.1, 143.9, 143.1, 142.3, 140.1, 136.9, 133.7, 130.0, 129.2(2×C), 127.7 (2×C), 123.2, 122.4, 120.6, 119.7, 112.9, 112.9, 109.8,55.5, 46.8, 36.0

3-[4-(2-benzimidazolylethyl)phenyl]phenol (VNPP355 (B19))

Method B: using VNPP319C (0.21 g, 0.7 mmol) and 3-hydroxyphenylboronicacid (0.15 g, 1.08 mmol). Yield (0.08 g, 37%): mp 226-227° C.; IR (Neat)2931, 1585, 1499, 1456, 1307, 1215, 1200, 785 cm⁻¹; ¹H NMR (500 MHz,DMSO-d₆) δ 3.16 (t, J=7.10 Hz, 2 H, CH₂), 4.52 (t, J=7.17 Hz, 2 H,═N—CH₂), 6.77 (d, J=7.48 Hz, 1 H, Ar-4³-H), 6.97-7.09 (m, 2 H, Ar-6³, 5¹Hs), 7.15-7.32 (m, 5 H, Ar-7¹, 2 ³, 5³, 2² and 6² Hs), 7.49 (d, J=7.78Hz, 2 H, Ar-3², 5² Hs), 7.59-7.73 (m, 2 H, Ar-8¹, 5 ¹ Hs), 8.10 (s, 1 H,Ar-2¹ H), 9.54 (s, 1 H, —OH); ¹³C NMR (500 MHz, DMSO-d₆) δ 158.4, 144.5,144.0, 141.9, 139.1, 138.0, 134.3, 130.5, 129.9 (2×C), 127.1 (2×C),122.8, 122.1, 120.0, 117.9, 114.9, 113.9, 111.1, 46.0, 35.6

3-[4-(2-benzimidazolylethyl)phenyl]benzenecarbonitrile (VNPP357 (B20))

Method B: using VNPP319C (0.2 g, 0.66 mmol) and 3-nitrilephenylboronicacid (0.15 g, 1 mmol). Yield (0.18 g, 83.5%): mp 157-159° C.; IR (Neat)3073, 2227, 1493, 1456, 1396, 1325, 1283, 1224, 796, 749 cm⁻¹; ¹H NMR(500 MHz, DMSO-d₆) δ 3.19 (t, J=7.32 Hz, 2 H, CH₂), 4.54 (t, J=7.32 Hz,2 H, ═N—CH₂), 7.20 (t, J=7.55 Hz, 1H, Ar-5¹ H), 7.26 (t, J=7.55 Hz, 1 H,Ar-6¹ H), 7.32 (d, J=8.09 Hz, 2H, Ar-2², 6² Hs), 7.63 (d, J=6.87 Hz, 1H, Ar-7¹ H), 7.64-7.70 (m, 4 H, Ar-3², 5², 4¹ and 5³ Hs), 7.80 (d,J=7.63 Hz, 1 H. Ar-6³ H), 8.00 (d, J=8.09 Hz, 1 H, Ar-4³-H), 8.09 (s, 1H, Ar-2³ H), 8.13 (s, 1H Ar-2¹ H); ¹³C NMR (500 MHz, CDCl₃) δ 144.1,143.1, 142.0, 138.0, 137.8, 133.6, 131.5, 130.9, 129.8, 129.7, 129.5,127.6, 127.6, 123.1, 122.3, 120.8, 120.6, 119.0, 113.1, 109.7, 46.6,36.0

1-(3′-methoxy-[1,1′-biphenyl]-4-yl)-1H-benzo[d]imidazole (VNPP444B(B23))

Method B: using VNPP442 and 3-methoxyphenylboronic acid. On purificationby FCC [petroleum ether/EtOAc (3:1)] gave solid VNPP444B(B23) (0.03 g,27%)¹H NMR (400 MHz, CDCl₃) δ 3.90 (s, 3 H, CH₃), 6.96 (dd, J=8.07, 2.20Hz, 1 H, Ar-4³-H), 7.18 (t, J=1.96 Hz, 1 H), 7.23 (d, J=7.58 Hz, 1 H),7.33-7.39 (m, 2 H), 7.39-7.45 (m, 1 H), 7.56-7.64 (m, 3 H), 7.79 (d,J=8.56 Hz, 2 H, Ar-3², 5²-Hs), 7.87-7.94 (m, 1 H), 8.16 (s, 1 H, Ar-2¹H); ¹³C NMR (101 MHz, CDCl₃) δ 160.13, 144.16, 142.25, 141.31, 140.95,135.60, 133.72, 130.05, 128.74, 124.28, 123.76, 122.86, 120.71, 119.61,113.15, 113.04, 110.51, 55.38

1-(3′-hydroxy-[1,1′-biphenyl]-3-yl)-1H-benzoimidazole (VNPP441C(B24))

Method B: using VNPP371/1 and 3-hydroxyphenylboronic acid. Onpurification by FCC [petroleum ether/EtOAc (3:1)] gave solidVNPP441C(B24) (0.047 g, 43%): ¹H NMR (400 MHz, CDCl₃) δ 6.98 (d, J=8.07Hz, 1 H, Ar-4³), 7.17 (br. s., 2 H), 7.20-7.24 (m, 1 H), 7.28-7.39 (m, 2H), 7.42 (d, J=7.34 Hz, 1 H), 7.49 (d, J=7.83 Hz, 1 H), 7.52 (s, 1 H),7.59 (t, J=7.83 Hz, 1 H), 7.68 (d, J=7.58 Hz, 1 H), 7.89 (d, J=7.82 Hz,1 H), 8.16 (s, 1 H, Ar-2¹); ¹³C NMR (101 MHz, CDCl₃) δ 157.58, 143.34,142.90, 141.93, 140.85, 136.36, 133.63, 130.44, 130.36, 127.06, 124.16,123.29, 122.89, 122.76, 120.19, 118.74, 115.77, 114.18, 110.75

1-(3′-methoxy-[1,1′-biphenyl]-3-yl)-1H-benzo[d]imidazole (VNPP420 (B25))

Method B: using VNPP371/1 and 3-methoxyphenylboronic acid. Onpurification by FCC [petroleum ether/EtOAc (3:1)] gave sticky solidVNPP420(B25) (0.1 g, 91%): ¹H NMR (400 MHz, CDCl₃) δ 3.88 (br. s., 3 H,CH₃), 6.96 (d, J=7.34 Hz, 1 H, Ar-4³ H), 7.15 (br. s., 1 H, Ar-6³ H),7.22 (br. s., 1 H, Ar-5¹ H), 7.35 (br. s., 2 H, Ar-6¹, 2³ Hs), 7.41 (br.s., 1 H, Ar-5³ H), 7.51 (br. s., 1 H, Ar-6² H), 7.59 (d, J=3.42 Hz, 1H,Ar 5² H), 7.64 (s, 1 H, Ar-4² H), 7.68 (s, 1 H, Ar-4¹ H), 7.72 (br. s.,1 H, Ar-7¹ H), 7.89 (br. s., 1 H, Ar-2² H), 8.17 (br. s., 1 H, Ar-2¹);¹³C NMR (101 MHz, CDCl₃) δ 160.1, 144.1, 143.3, 142.3, 141.2, 136.8,133.7, 130.4, 130.1, 126.8, 123.7, 122.8, 122.8, 122.8, 120.7, 119.6,113.4, 113.0, 110.5, 55.3; HRMS calcd 323.1154 (C₂₀H₁₆N₂O)Na⁺, found323.1156.

3-[4-(2-benzimidazolylethyl)phenyl]benzoicacid (VNPP359)

By slightly modifying Method B: using VNPP319C (0.2 g, 0.66 mmol) and3-carboxyphenylboronic acid (0.17 g, 1 mmol) and 2 M aq Na₂CO₃ (2×2.13mL). After completion of reaction, reaction mixture neutralized withdil. HCl, extracted with EtOAc, filtered and concentrated. Onpurification by FCC [petroleum ether/EtOAc (1:1) and then with 7%ethanol in DCM with traces of CH₃COOH] gave VNPP359 (0.07 g, 29%): mp215-217° C.; IR (Neat) 1689, 1505, 1438, 1310, 1242, 1164, 765, 754cm⁻¹; ¹H NMR (500 MHz, DMSO-d₆) δ 3.19 (t, J=7.25 Hz, 2 H, CH₂), 4.55(t, J=7.25 Hz, 2 H, ═N—CH₂), 7.19-7.24 (m, 1 H, Ar-5¹-H), 7.24-7.29 (m,1 H, Ar-6¹ H), 7.31 (d, J=8.09 Hz, 2 H, Ar-2², 6² Hs), 7.56-7.63 (m, 3H, Ar-3², 5², 7¹ Hs), 7.64-7.70 (m, 2 H, Ar-4¹, 5³ Hs), 7.86-7.92 (m, 1H, Ar-4³-H), 7.94 (d, J=7.78 Hz, 1 H, Ar-6³-H), 8.11 (s, 1 H, Ar-2¹ H),8.17 (m, 1 H, Ar-2³-H), 13.07 (br, 1 H, COOH); ¹³C NMR (500 MHz,DMSO-d₆) δ 167.9, 144.5, 144.0, 140.8, 138.6, 138.1, 134.3, 132.2,131.5, 130.1 (2×C), 129.9, 128.7, 127.7, 127.3 (2×C), 122.8, 122.1,120.0, 111.1, 45.9, 35.6

5-{3-[4-(benzimidazolylethyl)phenyl]phenyl}-1H-1,2,3,4-tetraazole(VNPP364)

Method C: using 0.85 g (0.26 mmol) of VPNN357. Yield (0.07 g, 76.5%): mp223-225° C.; IR (Neat) 1575, 1505, 1456, 1230, 992, 800, 742 cm⁻¹¹³; ¹HNMR (500 MHz, DMSO-d₆) δ 3 .20 (t, J=7.17 Hz, 3H, CH₂), 4.56 (t, J=7.25Hz, 2 H, ═N—CH₂), 7.20 (t, J=7.55 Hz, 1 H, Ar-5¹ H), 7.26 (t, J=7.48 Hz,1 H, Ar-6¹ H), 7.33 (d, J=7.93 Hz, 2 H, Ar-2², 6² Hs), 7.61-7.72 (m, 5H, Ar-3², 5², 6³, 5³ and 7¹ Hs), 7.86 (d, J=7.78 Hz, 1 H, Ar-4¹ H), 8.02(d, J=7.63 Hz, 1 H, Ar-4³ H), 8.10 (s, 1 H, Ar-2² H), 8.29 (s, 1 H,Ar-2⁴-H); ¹³C NMR (500 MHz, DMSO-d₆) δ 156.0, 144.5, 143.8, 141.4,138.8, 137.9, 134.3, 130.7, 130.2 (2×C), 129.7, 127.4 (2×C), 126.4,125.7, 125.6, 122.9, 122.1, 120.0, 111.2, 46.0, 35.6

6-(3-benzimidazolylphenyl)naphthalen-2-ol (VNPP372/2 (Na1))

Method B: using VNPP371/1 (0.1 g, 0.37 mmol) and6-hydroxynaphthyl-2-boronic acid (0.1 g, 0.53 mmol). On purification byFCC over short column [petroleum ether/EtOAc (1:1) and then with 2%ethanol in DCM with traces of TEA] gave VNPP372/2 (0.1 g, 81%): mp252-253° C.; IR (Neat) 1599, 1495, 1462, 1229, 1196, 962, 792, 741 cm⁻¹;¹H NMR (500 MHz, DMSO-d₆) δ 7.14 (d, J=8.54 Hz, 1 H, Ar-7³ H), 7.17 (d,J=2.5 Hz, 1 H, Ar-5³ H), 7.35 (p, J=7.4 Hz, 2 H, Ar-6¹ and 5¹ Hs), 7.67(d, J=7.8 Hz, 1 H, Ar-6² H), 7.70-7.75 (m, 2 H, Ar-4² and 5² Hs), 7.81(d, J=8.5 Hz, 2 H, Ar Hs), 7.87 (dd, J=8.7, 3.3 Hz, 2 H, Ar Hs), 7.90(d, J=7.6 Hz, 1 H, Ar-8³ H), 8.04 (s, 1 H, Ar-10³ H), 8.25 (s, 1 H,Ar-2¹ H), 8.70 (s, 1 H, Ar-2² H), 9.87 (s, 1H, —OH); ¹³C NMR (500 MHz,DMSO-d₆) δ 156.5, 144.5, 144.1, 142.9, 137.3, 134.8, 133.8, 131.2,130.6, 128.6, 127.4, 126.5, 126.4, 125.9, 124.2, 123.1, 122.7, 122.5,120.6, 119.8, 111.4, 109.1

6-(3-benzimidazolylphenyl)-2-methoxynaphthalene (VNPP373/2 (Na2))

Method B: using VNPP371/1 and 6-methoxynaphthyl-2-boronic acid Onpurification by FCC over short column [petroleum ether/EtOAc/TEA(4:2:0.1)] gave VNPP373/2 (Na2) (0.1 g, 81%): mp 156-157° C.; IR (Neat)1604, 1499, 1450, 1228, 1202, 1037, 837, 733 cm⁻¹; ¹H NMR (500 MHz,CDCl₃) δ 3.95 (br. s., 3 H, CH₃), 7.14-7.23 (m, 2 H), 7.36 (br. s., 2H), 7.51 (br. s., 1 H), 7.62 (br. s., 1 H), 7.67 (br. s., 1 H,), 7.73(d, J=8.56 Hz, 1 H), 7.80 (br. s., 2 H), 7.84 (br. s., 2 H), 7.90 (br.s., 1H), 8.02 (br. s., 1 H), 8.20 (br. s., 1 H); ¹H NMR (400 MHz,DMSO-d₆) δ 3.90 (br. s., 3 H), 7.38 (s, 2 H), 7.35 (s, 1 H), 7.73 (s, 1H), 7.71 (s, 2 H), 7.89-7.99 (m, 4 H), 8.07 (br. s., 1 H), 8.32 (br. s.,1 H), 8.71 (br. s., 1 H); ¹³C NMR (101 MHz, CDCl₃) δ 158.1, 144.1,143.4, 142.3, 136.9, 134.7, 134.2, 133.8, 130.4, 129.8, 129.1, 127.6,126.8, 125.9, 125.6, 123.7, 122.8, 122.7, 122.5, 120.7, 119.5, 110.5,105.6, 55.3; HRMS calcd 373.1311 (C₂₄H₁₈N₂O)H⁺, found 373.1314.

4-Hydrosystyrene (VNPP383/2)

To a solution of methyltriphenylphosphonium bromide (3.85 g, 10.8 mmol)in dry THF (30 mL), was added t-BuOK (1.3 g, 11.6 mmol) in severalportions over 30 min. Reaction mixture was stirred under argon for 1 h,then 4-hydroxybenzaldehyde (0.5 g, 4.09 mmol) was added, and stirringcontinued for another 3 h. The reaction mixture then diluted with DCM(100 ml), washed with water and brine, dried (Na₂SO₄) and solvents wereevaporated under vacuum. The crude product was purified by FCC[petroleum ether/EtOAc (4:1)] to get VNPP383/2 (0.25 g, 50.8%): mp101-105° C.; IR (Neat) 3299, 1611, 1597, 1509, 1443, 1365, 1222, 1171,824 cm⁻¹; ¹H NMR (500 MHz, CDCl₃) δ 4.13 (br, 1 H, —OH), 5.06-5.18 (m, 1H, vinyl CH₂—H), 5.54-5.65 (m, 1 H, vinyl CH₂ —H), 6.65 (dd, J=17.47,10.91 Hz, 1 H, Vinyl CH), 6.79 (dd, J=8.39 Hz, 2 H, Ar-2, 6 Hs), 7.30(dd, J=8.39 Hz, 2 H, Ar-3, 5 Hs)

4-[(1E)-2-(3-benzimidazolylphenyl)vinyl]phenol (VNPP388 (S1))

A mixture of p-Hydrosystyrene (VNPP383-2) (0.1 g, 0.83 mmol),1-(3-bromophenyl)-1H-benzo[d]imidazole (VNPP371/1) (0.12 g, 0.44 mmol),triethanolamine (0.065 g, 0.43 mmol) and Pd(II) acetate (0.004 g, 0.018mmol) was stirred under argon at 100° C. for 24 h. The reaction mixturewas cooled to room temperature, quenched by the addition of 2N HCl, andextracted with EtOAc (3×10 mL). the organic phases were dried (Na₂SO₄),evaporated and crude product was purified by FCC [petroleumether/EtOAc/TEA (3:2:0.01)] to get pure VNPP388 (0.025 g, 19%): mp182-184° C.; IR (Neat1596, 1580, 1496, 1454, 1278, 1232, 1217, 1171, 819cm⁻¹; ¹H NMR (400 MHz, DMSO-d6) δ 6.79 (m, J=8.31 Hz, 2 H, Ar-3³ and 3⁵Hs), 7.14 (d, J=16.38 Hz, 1 H, vinyl CH), 7.29-7.37 (m, 3 H, vinyl CH,Ar-1⁶ and 1⁵ Hs), 7.46 (m, J=8.31 Hz, 2 H, Ar-3² and 3⁶Hs), 7.53-7.66(m, 4 H, Ar-2⁶, 1⁴, 2⁵ and 2⁴ Hs), 7.79 (d, J=7.58 Hz, 1 H, Ar-1⁷ H),7.85 (s, 1 H, Ar-1² H), 8.60 (s, 1 H, Ar-2² H), 9.63 (s, 1 H, —OH); ¹³CNMR (400 MHz, DMSO-d6) δ 158.1, 144.3, 143.8, 140.2, 136.9, 133.6,130.7, 130.6, 128.6, 128.2, 125.7, 124.4, 123.9, 122.9, 122.4, 121.4,120.4, 116.0, 111.2

1-[(1E)-2-(3-benzimidazolylphenyl)vinyl]-4-methoxybenzene(VNPP391B3(S2))

Sythensized using P-Methoxyphenylstyren by following procedure forsynthesis of VNPP388 (S1). The crude product was purified by FCC[petroleum ether/EtOAc/DCM/TEA (6:1:1:0.01)] to get pure VNPP391B3(S2)(0.1 g, 83.4%): mp 95-96° C.; ¹H NMR (400 MHz, CDCl₃) δ 3.84 (s, 3 H,CH₃), 6.89-6.95 (m, 2 H, Ar-3³ and 5³ Hs), 7.03 (d, J=16.38 Hz, 1 H,vinyl CH), 7.15 (d, J=16.14 Hz, 1 H, vinyl CH), 7.33-7.40 (m, 3 H,Ar-6¹, 5¹ and 2³Hs), 7.46-7.51 (m, 2 H, Ar— 6³ and 6² Hs), 7.52-7.60 (m,3 H, Ar—, 4¹, 5² and 4² Hs), 7.62 (s, 1 H, Ar-2² H), 7.88-7.93 (m, 1 H,Ar-7¹ H), 8.16 (s, 1 H, Ar-2¹ H)¹³C NMR (400 MHz, CDCl₃) δ 159.7, 144.1,142.3, 139.9, 136.8, 133.7, 130.2, 130.2, 129.4, 128.0 (2×C), 125.8,124.9, 123.7, 122.8, 122.5, 121.5, 120.6, 114.2 (2×C), 110.5, 55.3; HRMScalcd 349.1311 (C₂₁H₁₈N₂O)Na⁺,—found 349.1314.

4-{[(3-benzimidazolylphenyl)methyl]amino}phenol (VNPP428(Am-a1))

Method D: using 4-hydroxybenaldehyde. Yield 33.8%; mp 94-97° C.; ¹H NMR(400 MHz, DMSO-d₆) δ 4.14-4.28 (m, 2 H, CH₂), 6.65-6.80 (m, 5 H), 7.19(d, J=7.34 Hz, 2 H), 7.26 (br. s., 2 H), 7.31 (br. s., 1 H), 7.74 (d,J=6.85 Hz, 1 H), 8.45 (br. s., 1 H), 9.32 (br. s., 1 H); ¹H NMR (400MHz, CDCl₃) δ 4.28 (s, 2 H, CH₂), 4.61 (s, 1 H, NH), 6.66 (t, J=2.08 Hz,1 H), 6.68-6.73 (m, 1 H), 6.75-6.80 (m, 1 H), 6.85-6.91 (m, 2 H),7.19-7.25 (m, 2 H), 7.27-7.32 (m, 2 H), 7.41 (d, J=4.16 Hz, 1 H),7.80-7.89 (m, 1 H), 8.09 (s, 1 H); ¹³C NMR (101 MHz, DMSO-d₆) δ 156.7,150.4, 143.4, 137.1, 130.7, 130.0, 128.9 (2×C), 128.2, 123.6, 122.7,120.3, 115.6 (2×C), 115.2, 112.4, 111.3, 110.6, 106.9, 46.3; HRMS calcd316.1444 (C₂₀H₁₇N₃O)H⁺, found 316.1446.

[(3-benzimidazolylphenyl)methyl](4-methoxyphenyl)amine (VNPP429(Am-a2))

Method D: using 4-methoxybenzaldehyde. Yield 50%; mp 121-22° C.; ¹H NMR(400 MHz, DMSO-d₆) δ 3.74 (br. s., 3 H), 4.22-4.34 (m, 2H), 6.63-6.79(m, 4 H), 6.93 (d, J=7.83 Hz, 2 H), 7.17-7.38 (m, 6H), 7.73 (d, J=6.85Hz, 1 H), 8.45 (br. s., 1 H); ¹³C NMR (101 MHz, CDCl₃) δ 159.0, 149.3,144.0, 142.3, 137.4, 133.6, 130.6, 130.4, 128.7 (2×C), 123.4, 122.6,120.4, 114.2 (2×C), 112.6, 112.4, 110.8, 107.6, 55.3, 47.5; HRMS calcd352.1420 (C₂₁H₁₉N₃O)Na⁺, found 352.1422.

N-(3-benzimidazolylphenyl)(4-methoxyphenyl)carboxamide (VNPP432(Amd-b2))

A mixture of VNPP423 (0.15 g, 0.72 mmol), TEA (0.1 g, 1.0 mmol) and4-methoxybenzoyl chloride (0.3 g, 1.8 mmol) in EtOAc (10 mL) stirred atroom temperature for 5 hrs. Reaction mixture filtered, filtrate washedwith pet ether-EtOAc mixture (3:2), then with water. Mother liquorpurified separately by FCC [1% ethanol in DCM with traces of TEA].Overall yield 0.12 g (48.7%): mp 212-214° C.; ¹H NMR (400 MHz, CDCl₃) δ3.83 (s, 2 H), 6.93 (d, J=6.85 Hz, 2 H) 7.06 (d, J=7.82 Hz, 1 H) 7.19(t, J=7.58 Hz, 1 H) 7.31-7.44 (m, 4 H) 7.77-7.81 (m, 2 H) 7.97 (d,J=8.31 Hz, 1 H) 8.02 (s, 1 H) 8.31 (s, 1 H, Ar-2¹) 8.72 (s, 1 H, NH);HRMS calcd 344.1393 (C₂₁H₁₇N₃O₂)H⁺, found 344.1397.

(3-benzimidazolylphenyl)[(4-methoxyphenyl)sulfonyl]amine(VNPP431(SulAmd-2))

A mixture of VNPP423 (0.15 g, 0.72 mmol), 4-methoxybenzenesulfonylchloride (0.44 g, 2.15 mmol) in pyridine was refluxed for 24 h. Cooledto room temperature, poured on ice cold water, extracted with EtOAc,evaporated and crude product was purified by FCC [1% ethanol in DCM withtraces of TEA]. VNPP431(SulAmd-2) (0.13 g, 47.8%): mp 169-71° C.; ¹H NMR(400 MHz, CDCl₃) δ 3.84 (s, 3 H, CH₃) 6.90-6.97 (m, 2 H, Ar-2², 4² Hs)7.13-7.19 (m, 1 H, Ar-6² H) 7.21-7.26 (m, 1 H, Ar-5² H) 7.28-7.31 (m, 2H, Ar—Hs) 7.32 (d, J=1.96 Hz, 1 H, Ar—H) 7.33-7.40 (m, 2 H, Ar—Hs)7.41-7.48 (m, 1 H, Ar—H) 7.76-7.81 (m, 2 H, Ar—Hs) 7.83-7.88 (m, 1 H,Ar—H) 8.07 (s, 1 H, Ar-2¹); ¹³C NMR (101 MHz, CDCl₃) δ 163.4, 143.9,142.0, 138.6, 137.2, 133.3, 131.0, 130.4, 129.5 (2×C), 123.9, 123.0,120.6, 120.1, 120.0, 115.9, 114.4 (2×C), 110.4, 55.6; HRMS calcd402.0882 (C₂₀H₁₇N₃O₃S)Na⁺, found 402.0886.

6β-imidazol-1yl-3,5-cycloandrostan-17-benzimidazole-1-yl-16-ene(VNPP433-6β), 3α-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene (VNPP433-3α(A3)) and3β-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl) androsta-5,16-diene(VNPP433-3β(2d)(A1))

A round bottom flask equipped with Dean-stark apparatus and condenser ischarged with VNPT88 (0.6 g, 1.3 mmol), imidazole (0.3 g, 4.4 mmol) andtoluene. Reaction mixture refluxed for 12 h. Reaction mixture cooled,solvent evaporated under vacuum, residue washed with water to get crudeproduct. Purification by FCC [1% ethanol in DCM with traces of TEA] toget product of three components VNPP433-6β, VNPP433-3α(A3) andVNPP433-3β(2d)(A1) (0.23 g, 61%): This three component mixture subjectedto preparative HPLC for the isolation of individual compounds.

HPLC separation: Preparative HPLC separation was performed on WatersPrep Nova-Pak (7.8×300 mm, 60 Å, 6 μm) HR C-18 reversed-phase HPLCcolumn (Waters, Milford, Mass.) coupled with Waters 2489 UV/visibledetector operated at 254 nm. Elution was performed using Waters model2535 Quarternary Gradient Module pump to deliver a constant flow rate of6 mL/min. The solvent system consisted of Water/MeOH/CH₃CN (200:500:300,v/v/v+500 μL of TEA per 1000 mL of mobile phase) and maintainedisocratically. Sample stock solution was prepared by dissolving 200 mg(three component mixture obtained by FCC) in 20 mL of mobile phase.Total of twenty injections each 1 mL volume with run time of 13 minutesperformed. Compound VNPP433-6β was collected between 5.12-6.73 min,VNPP433-3α(A3) collected between 7.77-8.93 min and VNPP433-3β(2d)(A1)was collected between 10.07-12.09 min.

Example of HPLC chromatogram of three component mixture. The retentiontimes for VNPP433-6β, VNPP433-3α(A3) and VNPP433-3β(A1) were 5.14, 7.87and 10.07 min, respectively. See FIG. 11.

6β-imidazol-1yl-3,5-cycloandrostan-17-benzimidazole-1-yl-16-ene(VNPP433-6β)

Retension time 5.14 min; 0.13 g, 34.6%, mp 104-106° C.; ¹H NMR (400 MHz,CDCl₃) δ 0.79 (s, 3 H, 18-CH₃), 0.97 (s, 3 H, 19-CH₃), 3.66 (br. s., 1H, 6α-H), 5.97-6.02 (m, 1 H, 16-H), 7.09 (s, 1 H, Ar-4²-H), 7.22 (br.s., 1 H, Ar-5²-H), 7.28-7.36 (m, 2 H, Ar-5¹, 6¹-Hs), 7.46-7.52 (m, 1 H,Ar-7¹-H), 7.79-7.89 (m, 2 H, Ar-4¹-H and Ar-2²-H), 7.95 (s, 1 H,Ar-2¹-H); ¹H NMR (400 MHz, DMSO-d₆) δ ppm 0.67 (s, 3 H), 0.91 (s, 3H),3.73 (br. s., 1 H), 6.09 (s, 1 H), 6.93 (br. s., 1 H), 7.21-7.33 (m,2H), 7.36 (s, 1 H), 7.57 (d, J=7.82 Hz, 1 H), 7.70 (d, J=7.58 Hz, 1 H),7.85 (br. s., 1 H), 8.25 (s, 1 H); ¹³C NMR (400 MHz, CDCl₃)δ147.35,143.24, 141.49, 136.41, 134.47, 128.62, 123.52, 123.48, 122.54, 120.21,118.13, 111.07, 58.61, 55.37, 48.47, 47.42, 43.24, 35.18, 34.90, 34.34,33.34, 30.14, 29.19, 24.78, 24.24, 22.01, 19.72, 16.25, 14.31; HRMScalcd 439.2856 (C₂₉H₃₄N₄O)H⁺, found 439.2858.

3α-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl) androsta-5,16-diene(VNPP433-3α(A3))

Retention time 8.87 min; 0.012 g, 3.2%: mp 199-200° C.; ¹H NMR (400 MHz,CDCl₃) δ 1.02 (s, 3 H, 18-CH₃), 1.14 (s, 3 H, 19-CH₃), 4.41 (br. s., 1H, 3δ-H), 5.56 (br. s., 1 H, 6-H), 5.99 (br. s., 1 H, 16-H), 7.03 (d,J=4.89 Hz, 2 H, Ar-4²-H and Ar-5²-H), 7.28-7.33 (m, 2 H, Ar-5¹, 6¹-Hs),7.49 (d, J=8.31 Hz, 1 H, Ar-7¹-H), 7.73 (s, 1 H, Ar-2²-H), 7.81 (d,J=8.80 Hz, 1 H, Ar-4¹-H), 7.95 (s, 1 H, Ar-2¹-H); ¹H NMR (400 MHz,DMSO-d₆) δ 0.97 (s, 3 H, 18-CH₃), 1.10 (s, 3 H, 19-CH₃), 4.42 (br. s., 1H, 3δ-H), 5.53 (br. s., 1 H, 6-H), 6.06 (br. s., 1 H, 16-H), 6.86 (s, 1H), 7.19-7.33 (m, 3 H), 7.56 (d, J=7.83 Hz, 1 H), 7.69 (d, J=7.34 Hz, 2H), 8.24 (s, 1 H); ¹³C NMR (400 MHz, CDCl₃) δ 147.11, 143.29, 141.64,139.13, 134.58, 128.56, 124.18, 123.41, 123.36, 122.48, 120.22, 118.70,111.13, 55.74, 53.00, 50.19, 47.23, 37.27, 35.97, 34.74, 32.29, 31.12,30.23, 30.20, 29.71, 28.39, 20.27, 19.32, 16.01; HRMS calcd 439.2856(C₂₉H₃₄N₄O)H⁺, found 439.2857

3β-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl) androsta-5,16-diene(VNPP433-3β(2d)(A1))

Retention time 10.074 min; 0.04 g, 10.6%: mp 198-200° C.; 1H NMR (400MHz, CDCl3) δ 1.04 (s, 3 H, 18-CH3), 1.14 (s, 3 H, 19-CH3), 3.86-3.98(m, 1 H, 3α-H), 5.50 (br. s., 1 H, 6-H), 6.00 (br. s., 1 H, 16-H), 7.00(br. s., 1 H, Ar-42-H), 7.07 (br. s., 1 H, Ar-52-H), 7.28-7.33 (m, 2 H,Ar-51, 61-Hs), 7.49 (d, J=5.62 Hz, 1 H, Ar-71-H), 7.57 (br. s., 1 H,Ar-22-H), 7.82 (d, J=5.62 Hz, 1 H, Ar-41-H), 7.96 (s, 1 H, Ar-21-H); 1HNMR (400 MHz, DMSO-d6) δ ppm 0.99 (s, 3 H), 1.12 (s, 3 H), 3.93-4.03 (m,1 H), 5.47 (d, J=4.65 Hz, 1 H), 6.07 (s, 1 H), 6.88 (s, 1 H), 7.23-7.32(m, 2 H), 7.33 (s, 1 H), 7.57 (d, J=7.58 Hz, 1 H), 7.71 (d, J=7.58 Hz, 1H), 7.74 (s, 1 H), 8.27 (s, 1 H) 13C NMR (400 MHz, CDCl3) δ 147.25,143.40, 141.72, 140.20, 134.68, 129.27, 124.21, 123.55, 122.63, 122.39,120.34, 116.97, 111.26, 57.54, 55.92, 50.62, 47.35, 40.67, 38.00, 37.03,34.94, 31.17, 30.41, 30.02, 29.83, 20.73, 19.47, 16.15, 0.14; HRMS calcdcalcd 439.2856 (C₂₉H₃₄N₄O)H+, found 439.2858.

3β-(1H-imidazole-1-carbothioate)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene (VNPP397 (A9))

A solution of 17-(1H-benzimidazol-1-yl) androsta-5,16-dien-3β-ol(VN/124-1) (0.2 g, 0.515 mmol), 1,1′-thiocarbonyldiimidazole (0.18 g,1.03 mmol) in anhydrous acetonitrile (2 mL) and DCM (1 mL) was refluxedfor 5 h. The solvent evaporated, residue treated with water, andfiltered. The crude brown solids obtained was purified by FCC using 1.7%ethanol in DCM in presence of traces of TEA (0.06%) to give VNPP397(0.14 g, 54.5%): mp 187-88° C.; IR (Neat) 1487, 1456, 1386, 1324, 1291,1243, 1222, 1110, 986, 827, 743 cm¹ ¹H NMR (400 MHz, CDCl₃) □ 1.03 (s, 3H, 18-CH₃), 1.14 (s, 3 H, 19-CH₃), 5.31-5.42 (m, 1 H, 3α-H), 5.48-5.57(m, 1 H, 6-H), 6.00 (br. s., 1 H, 16-H), 7.03 (s, 1 H, Ar-4² H),7.28-7.35 (m, 2 H, Ar-5¹ and 6¹ Hs), 7.49 (d, J=5.14 Hz, 1 H, Ar-5² H),7.64 (s, 1 H, Ar-4¹ H), 7.82 (d, J=7.58 Hz, 1 H, Ar-7¹ H), 7.96 (s, 1 H,Ar-2¹), 8.35 (s, 1 H, Ar-2² H); ¹³C NMR (400 MHz CDCl₃) □183.3, 147.1,143.3, 141.6, 139.0, 136.7, 130.7, 124.1, 123.4, 123.2, 122.5, 120.2,117.9, 111.1, 83.3, 55.7, 50.3, 47.2, 37.2, 36.8, 36.6, 34.8, 31.1,30.3, 30.2, 27.0, 20.6, 19.2, 16.0

3β-(pyridine-3-ylmethoxy)-17-(1H-benzimidazol-1-yl) androsta-5,16-diene(VNPP415C (A15))

To a solution of VN124-1 (0.1 g, 0.257 mmol), Sodium hydride (0.02 g,0.836 mmol) and DMF (2 mL) at room temperature under argon added3-(iodomethyl) pyridine hydroiodide (0.16 g, 0.46 mmol). The reactionheated to 65° C., continued for 48 hr, cooled and then poured onto icecold water, filtered and dried. Purification by FCC [petroleumether/EtOAc (3:2)] gave VNPP415C(A15) (53 mg, 43%): mp 196-97° C.; ¹HNMR (400 MHz, CDCl₃) δ 1.02 (s, 3 H, 18-CH₃), 1.07 (s, 3 H, 19-CH₃),3.31 (m, 1 H, 3α-H), 4.59 (s, 2 H, —CH₂), 5.42 (d, J=5.38 Hz, 1 H, 6-H),5.98 (dd, J=3.18, 1.71 Hz, 1 H, 16-H), 7.27-7.34 (m, 3 H, Ar-Hs),7.47-7.51 (m, 1 H), 7.70 (d, J=7.82 Hz, 1 H), 7.78-7.85 (m, 1 H), 7.96(s, 1 H, Ar-2¹ H), 8.54 (dd, J=5.01, 1.59 Hz, 1 H, Ar-6² H), 8.59 (d,J=1.71 Hz, 1 H, Ar-2² H); ¹³C NMR (101 MHz, CDCl₃) δ 149.14, 149.01,147.23, 143.33, 141.64, 141.13, 135.28, 134.58, 134.37, 124.06, 123.39,122.46, 121.19, 120.23, 111.14, 97.23, 78.86, 67.50, 55.90, 50.56,47.25, 39.11, 37.11, 37.09, 34.89, 31.14, 30.38, 30.30, 28.34, 20.67,19.32, 16.02

3β-(pyridine-4-ylmethoxy)-17-(1H-benzimidazol-1-yl) androsta-5,16-diene(VNPP414(A16))

To a solution of VN124-1 (0.1 g, 0.257 mmol), Sodium hydride (0.185 g,7.7 mmol) and DMF (2 mL) at room temperature under argon added4-(bromomethyl)pyridine hydrobromide (0.2 g, 0.772 mmol). The reactionheated to 65° C., continued for 12 hr, cooled and then poured onto icecold water, filtered and dried. Purification by FCC [petroleumether/EtOAc (3:2)] gave VNPP414(A16) (15 mg, 12%): mp 166-67° C.; ¹H NMR(400 MHz, CDCl₃) δ 1.02 (s, 3 H, 18-CH₃), 1.08 (s, 3 H, 19-CH₃), 3.30(m, 1 H, 3α-H), 4.59 (s, 2 H, —CH₂), 5.42 (d, J=4.89 Hz, 1 H, 6-H), 5.98(br. s., 1 H, 16-H), 7.27-7.34 (m, 4 H, aromatic-Hs), 7.49 (d, J=5.14Hz, 1 H, Ar7¹-H), 7.81 (d, J=4.40 Hz, 1 H, Ar-6¹-H), 7.95 (s, 1 H,Ar-2¹-H), 8.57 (d, J=4.65 Hz, 2 H, Ar-2², 6²-Hs); ¹³C NMR (101 MHz,CDCl₃) δ 149.8, 148.2, 141.6, 141.0, 134.5, 124.0, 123.3, 122.4, 121.7,121.2, 120.2, 111.1, 79.1, 68.3, 55.9, 50.5, 47.2, 39.1, 37.1, 37.0,34.8, 31.1, 30.3, 30.3, 28.3, 20.6, 19.3, 16.0; HRMS calcd 502.2828(C₃₂H₃₇N₃O)Na⁺, found 502.2834.

3β-(methoxy)-17-(1H-benzimidazol-1-yl) androsta-5,16-diene(VNPP412(A31))

To a solution VN124-1 (0.1 g, 0.257 mmol), Sodium hydride (12.4 mg,0.515 mmol) and DMF (2 mL) at room temperature under argon added MeI (75mg, 0.515 mmol). The reaction continued for 1 hr and then poured ontoice cold water, extracted with EtOAc and organic solvent evaporated. Thecrude product was purified by FCC [petroleum ether/EtOAc/TEA (4:1:0.2)]to give VNPP412(A31) (0.08 g, 77%): mp 136-37° C.; ¹H NMR (400 MHz,CDCl₃) δ 1.02 (s, 3 H, 18-CH₃), 1.05 (s, 3 H, 19-CH₃), 2.43 (d, J=13.69Hz, 2 H), 3.08 (m, 1 H, 3α-H), 3.37 (s, 3 H, CH₃), 5.41 (br. s., 1 H,6-H), 5.98 (br. s., 1 H, 16-H), 7.30 (m, 2 H, Ar-5¹, 6¹-Hs), 7.49 (d,J=3.34 Hz, 1 H, Ar-7¹-H), 7.81 (m, 1 H, Ar-4¹-H), 7.96 (s., 1 H,Ar-2¹-H); ¹³C NMR (101 MHz, CDCl₃) δ 147.2, 141.3, 124.0, 123.3, 122.4,120.9, 120.2, 111.1, 80.1, 55.9, 55.6, 50.5, 47.2, 38.7, 37.1, 37.0,34.9, 31.1, 30.3, 30.3, 27.9, 20.6, 19.3, 16.0; HRMS calcd 425.2563(C₂₇H₃₄N₂O)Na⁺, found 425.2566.

3-[{Trifluoromethyl-sulfonyl}-oxy]-estra-1,3,5-(10)-trien-17-one(VNPP308)

To estrone (2.5 g, 9.24 mmol) in DCM (40 mL) at 0° C. was added TEA (1.9g, 18.5 mmol) and truffle anhydride (2.86 g, 10.17 mmol,). The reactionmixture was stirred at 0° C. for 1 h before the addition of saturatedaqueous NaHCO₃ (40 mL). The phases were separated and the aqueous phasewas extracted with DCM (2×40 mL). The combined organic phases are washedwith brine (40 mL) and dried (Na₂SO₄). The filtrate was concentrated invacuo and the residue was purified by short FCC [petroium ether/EtOAc(8:2)] to give VNPP308 (1.67 g, 99%): mp 99° C.; IR (Neat) 1736, 1487,1420, 1209, 1138, 916, 829 c m-1; ¹H NMR (400 MHz, CDCl3) δ 0.92 (s, 3H, 18-CH₃), 1.43-1.71 (m, 7 H), 1.95-2.22 (m, 4 H), 2.30 (td, J=10.60,4.39 Hz, 1 H, 12β-H), 2.37-2.45 (m, 1 H, 14-H), 2.48-2.58 (m, 1 H, 8-H),2.95 (dd, J=8.66, 3.89 Hz, 2 H, 6-H), 7.00 (d, J=2.76 Hz, 1 H, 4-H),7.03 (dd, J=8.64, 2.68 Hz, 1 H, 2-H), 7.34 (d, J=8.28 Hz, 1 H, 1-H

Estra-1,3,5-(10)-trien-17-one-3-methylcarboxylate (VNPP309A)

A mixture of VNPP308 (3 g, 7.46 mmol), Pd(II) acetate (0.12 g, 0.54mmol), 1,3-bis-(diphenyl-phosphino)propane (dppp) (0.19 g, 0.46 mmol),TEA (2.36 g, 23.27 mmol), methanol (12 mL), and DMF (20 mL) were heatedat 70° C. with a slow constant purging of carbon monoxide for 9 h. Thereaction mixture was then cooled, poured into brine (100 mL), andextracted with EtOAc (3×100 mL), filtered through celite and dried withNa₂SO₄ and concentrated under vacuum to give crude product, which wasthen purified by FCC [petroleum ether/EtOAc (9.5:0.5, 9:1, 8.5:1.5gradient)] to give first pure VNPP309A (0.75 g, 32.2%) mp. 120-122° C.;IR (Neat) 2931, 1731, 1717, 1441, 1290, 1260, 1199, 1174, 750 c m-1¹HNMR (400 MHz, CDCl3) δ 0.92 (s, 3 H, 18-CH₃), 1.43-1.70 (m, 8H), 1.99(d, J=12.05 Hz, 1 H,), 2.04-2.21 (m, 3 H), 2.47-2.57 (m, 1 H, 8-H),2.94-3.00 (m, 2 H, 6-H), 3.90 (s, 3 H, CH₃), 7.36 (d, J=8.03 Hz, 1 H,1-H), 7.78 (d, J=1.6 Hz, 1 H, 4-H), 7.80 (m, 1 H, 2-H)

Then UV inactive compound (Estrone) (1.2 g): mp 255-262° C.: IR (Neat)3281, 1706, 1578, 1496, 1286, 1246, 1152, 1054, 815; ¹H NMR (500 MHz,CDCl₃) δ 0.91 (s, 3H. 18-CH₃), 1.41-1.65 (m, 8 H), 1.93-2.08 (m, 3 H),2.10-2.18 (m, 1 H), 2.21-2.27 (m, 1 H), 2.35-2.41 (m, 1 H,), 2.50 (dd,J=19.00, 8.62 Hz, 1 H), 2.87 (dd, J=6.56, 3.05 Hz, 2 H, 6-H), 4.62 (s, 1H, —OH), 6.58 (d, J=2.3 Hz, 1 H, H-4), 6.64 (dd, J=8.32, 2.37 Hz, 1 H,H-2), 7.15 (d, J=8.24 Hz, 1 H, H-1); ¹H NMR (400 MHz, DMSO-d6) δ 0.82(s, 3 H), 1.31-1.40 (m, 3 H), 1.43-1.52 (m, 2 H), 1.56 (br. s., 1 H),1.75 (d, J=8.56 Hz, 1 H), 1.94 (d, J=8.07 Hz, 2 H), 2.31 (d, J=5.14 Hz,1 H), 2.75 (d, J=5.14 Hz, 2 H), 6.45 (s, 1 H), 6.51 (d, J=8.07 Hz, 1 H),7.05 (d, J=8.56 Hz, 1 H), 9.00 (s, 1 H)

Estra-1,3,5-(10),16-tetraen-17-bromo-16-formyl-3-methylcarboxylate(VNPP315B)

To ice cold DMF (2.5 mL) drop wise added phosphorus tribomide (1.44 g,5.30 mmol) and stirred for 5 min. A solution of VNPP309A (0.75 g, 2.40mmol) in dry CHCl₃ (5 mL) was added and refluxed for 5 h. It was thenconcentrated under vacuum, poured on to ice, extracted with EtOAc,organic phase dried and evaporated to get solid. Residue then moistenedwith THF, stirred with 20% HCl for 2 h, extracted with EtOAc, organicphase dried with Na₂SO₄ and evaporated. On purification by FCC[petroleum ether/EtOAc (9:1)] gave pure VNPP315B (0.5 g, 51.7%): mp.149-150° C.; IR (Neat) 2936, 1708, 1668, 1578, 1435, 1291, 1260, 1193,1108, 757 c m-1; ¹H NMR (400 MHz, CDCl3) δ 0.97 (s, 3 H, 18-CH₃), 1.55(s, 1 H), 1.64 (d, J=1.25 Hz, 1 H), 1.65-1.68 (m, 1 H), 1.68-1.71 (m, 1H), 1.71-1.81 (m, 1 H), 1.96-2.04 (m, 2 H), 2.09-2.18 (m, 1 H), 2.39(br. s., 1 H), 2.47-2.55 (m, 1 H), 2.65 (dd, J=14.68, 6.40 Hz, 1 H),2.93-3.00 (m, 2 H, 6-H), 3.90 (s, 3 H, CH₃), 7.34 (dd, J=8.28, 1.0 Hz,1H, 1-H), 7.78 (d, J=1.7 Hz,1 H, 4-H), 7.80 (dd, J=8.16, 1.88 Hz, 1 H,2-H), 9.91 (s, 1 H, CHO)

Estra-1,3,5-(10),16-tetraen-17-(1H-benzimidazol-1-yl)-16-formyl-3-methylcarboxylate(VNPP330)

A mixture of compound VNPP315B (0.25 g, 0.62 mmol), benzimidazole (0.22g, 1.86 mmol), and K₂CO₃ (0.26 g, 1.86 mmol) in dry DMF (2 mL) washeated at 80° C. under argon for 1 h. After cooling to room temperature,the reaction mixture was poured onto ice-cold water (50 mL), and theresulting precipitate was filtered, washed with water, and dried. Onpurification over short FCC [hexane/EtOAc/TEA (10:5:0.1)] gave pureVNPP330 (0.26 g, 95%): mp 121-122° C.: IR (Neat) 2945, 1705, 1664, 1602,1489, 1291, 1086, 748; ¹H NMR (500 MHz, CDCl3) δ 1.02 (br. s., 3 H,18-CH₃), 1.18-1.32 (m, 1 H), 1.53-1.64 (m, 4 H), 1.67 (d, J=9.61 Hz, 1H), 1.76 (d, J=10.99 Hz, 2 H), 2.00 (br. s., 2 H), 2.04 (br. s., 1 H),2.06-2.14 (m, 1 H), 2.38-2.61 (m, 3H), 2.88 (s, 3 H), 2.96 (s, 2 H),2.98-3.08 (m, 2 H), 3.90 (s, 3 H, CH₃), 7.31-7.42 (m, 4 H, Ar—H), 7.80(br. s., 2 H, Ar—H), 7.88 (dd, J=6.26, 2.90 Hz, 1 H, 2-H), 8.02 (br. s.,1 H, Ar-2¹-H), 9.62 (s, 1 H, CHO)

3-(methylcarboxylate)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien(VNPP334 (E6))

A solution of VNPP330 (0.22 g, 0.5 mmol) in dry benzonitrile (3 mL) wasrefluxed in the presence of 10% Pd on activated charcoal (0.11 g, i.e.,50% weight of the substrate) for 9 h. Then 0.05 g more catalyst addedand refluxed for 12 h. After cooling to room temperature, the catalystwas removed by filtration through a Celite pad. The filtrate wasevaporated, and the residue was purified by FCC [petroleumether/EtOAc/TEA (8:2:0.5)] to give VNPP334 (0.12 g, 57%): mp: 165-167°C.: IR (Neat) 2927, 1716, 1486, 1451, 1288, 1265, 1108, 753, 737; ¹H NMR(500 MHz, CDCl3) δ 1.03 (s, 3 H, 18-CH₃), 1.19-1.32 (m, 1 H), 1.53-1.61(m, 1 H), 1.63-1.81 (m, 3 H), 1.87 (td, J=12.36, 3.20 Hz, 1 H),1.93-2.12 (m, 3 H), 2.25-2.36 (m, 1 H), 2.42-2.59 (m, 3 H), 2.95-3.06(m, 2 H, 6-Hs), 3.90 (s, 3 H, CH₃), 6.02 (br. s., 1 H, 16-H), 7.28-7.39(m, 3 H, Ar—Hs), 7.51 (d, J=6.41 Hz, 1 H, Ar-7¹ H), 7.77-7.86 (m, 3 H,Ar—Hs), 8.00 (s, 1 H, Ar-2¹ H); CDCl3: 167.46, 147.38, 145.47, 143.44,141.79, 136.91, 134.77, 130.42, 127.82, 127.06, 125.29 124.28, 123.68,122.76, 120.43, 111.30, 55.31, 52.19, 47.76, 45.06, 36.82, 35.07, 30.23,29.32, 27.22, 26.11, 16.37

3-(carboxylic acid)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien(VNPP341 (E5))

A suspension of VNPP334 (0.1 g, 0.24 mmol), 10% methanolic KOH (1.5 mL)and methanol (5 mL) refluxed for 2 h. Reaction mixture cooled, solidfiltered and washed with cold methanol. On purification by FCC overshort column [10% methanol in DCM] gave pure VNPP341 (0.09 g, 93%): mp270° C. (decomp): IR (Neat) 1693, 1606, 1494, 1456, 1299, 1252, 1226,1122, 756, 745, 737; ¹H NMR (500 MHz, DMSO-d₆) δ 0.96 (s, 3 H, CH₃),1.19-1.29 (m, 1 H), 1.42-1.58 (m, 2 H), 1.60-1.72 (m, 1 H), 1.76-1.90(m, 2 H), 1.90-2.02 (m, 2 H), 2.36-2.49 (m, 3 H), 2.93 (d, J=5.34 Hz, 2H, 6-H), 6.08 (br. s., 1 H, 16-H), 7.23-7.34 (m, 2 H, Ar—Hs), 7.37 (d,J=8.24 Hz, 1 H, 1-H), 7.59 (d, J=7.93 Hz, 1 H, Ar-4¹ H), 7.65-7.77 (m, 3H), 8.30 (s, 1 H, Ar-2¹ H); ¹³C NMR (500 MHz, DMSO-d₆) δ 168.2, 147.2,145.5, 143.5, 142.9, 137.1, 134.9, 130.4, 127.2, 125.7, 124.4, 123.9,122.8, 120.2, 112.0, 55.1, 47.5, 44.8, 36.7, 34.5, 30.2, 29.2, 27.1,26.1, 19.5, 16.4; ¹H NMR (500 MHz, CDCl3) δ 1.06 (s, 3 H), 1.57-1.66 (m,2 H), 1.67-1.81 (m, 3H), 1.85-1.96 (m, 2 H), 1.96-2.16 (m, 5 H),2.29-2.39 (m, 2 H), 2.46-2.62 (m, 4 H), 3.04 (d, J=4.88 Hz, 2 H), 6.06(br. s., 1 H), 7.32-7.37 (m, 2 H), 7.39 (d, J=8.09 Hz, 1 H), 7.51-7.58(m, 1 H), 7.84-7.94 (m, 3 H), 8.06 (s, 1 H)

3-(Acetoxy)-estra-1,3,5-(10)-trien-17-one (VNPP310)

To a ice cold solution of estrone (3 g, 11.1 mmol) in pyridine (15 mL)added acetic anhydride (7.36 g, 72.1 mmol) and stirred at roomtemperature 16 h. Reaction mixture poured on ice water mixture (300 mL)and resulting solid filtered, washed with water and dried to gave pureVNPP310 (3.4 g, 98%): mp 222-223° C.; IR (Neat) 1759, 1732, 1491, 1365,1210, 1183, 1019, 821; ¹H NMR (300 MHz, CDCl₃) δ 0.89 (s, 3 H, 18-CH₃),2.29 (s, 3 H, OAc), 2.85-2.96 (m, 2 H), 6.81 (m, 1 H, 1-H), 6.85 (d,J=8.78 Hz, 1 H, H-2), 7.29 (s, 1 H, H-4)

3-(Acetoxy)-17-(bromo)-16-(formyl)-estra-1,3,5-(10)-trien (VNPP311)

To ice cold DMF (6.5 mL) drop wise added phosphorus tribromide (2.88 g,10.63 mmol) and stirred for 5 min. To this, a solution of VNPP310 (1.5g, 5.4 mmol) in dry CHCl₃ (20 mL) was added and refluxed for 5 h. It wasthen concentrated under vacuum, poured on to ice, extracted with EtOAc,organic phase dried and evaporated to get solid. Residue then moistenedwith THF (˜2 mL), stirred with 20% HCl (15 mL) for 2 h, extracted withEtOAc, organic phase dried with Na₂SO₄ and evaporated. On purificationby FCC [petroleum ether/EtOAc (8:2)] gave pure VNPP311 (0.5 g, 26%): mp123-25° C.; 1765, 1670, 1584, 1493, 1366, 1189, 1013, 897; ¹H NMR (500MHz, CDCl3) δ 0.96 (s, 3 H, 18-CH₃), 1.40-1.52 (m, 1 H), 1.57-1.70 (m, 3H), 1.92-2.02 (m, 2 H), 2.12 (dd, J=14.72, 11.67 Hz, 1 H), 2.29 (s, 3 H,OAc), 2.46 (d, J=9.46 Hz, 1 H), 2.63 (dd, J=14.72, 6.33 Hz, 1 H),2.85-2.96 (m, 2 H, C-6 Hs), 6.80-6.83 (m, 1 H, 1-H), 6.86 (dd, J=8.47,2.37 Hz, 1 H, 2-H), 7.27 (s, 1 H, 4-H), 9.91 (s, 1 H, CHO); C₂₁H₂₃BrO₃,402.083

Estra-1,3,5-(10),16-tetraen-17-bromo-16-formyl-3-ol (VNPP312)

A solution of VNPP311 (0.4 g, 1 mmol), 10% ethanolic KOH (2 mL) andethanol (5 mL) stirred at room temperature over night. Reaction mixtureevaporated, residue treated with cold water, filtered. On purificationby FCC [petrolium ether/EtOAc (1:1)] gave (0.1 g, 28%): mp 149-151° C.;IR (Neat) 1718, 1609, 1462, 1497, 1287, 1250 1100, 837, 781; ¹H NMR (500MHz, CDCl3) δ 0.96 (s, 3 H, 18-CH₃), 1.38-1.49 (m, 1 H), 1.57-1.68 (m, 3H), 1.75 (td, J=11.53, 6.59 Hz, 1 H), 1.90-2.02 (m, 2 H), 2.07-2.16 (m,1 H), 2.29 (d, J=5.49 Hz, 1 H), 2.39-2.49 (m, 1 H), 2.63 (dd, J=15.37,6.59 Hz, 1 H), 2.81-2.95 (m, 2 H, 6-H), 6.58 (s, 1 H, 4-H), 6.64 (d,J=8.78 Hz, 1 H, H-2), 7.13 (d, J=8.78 Hz, 1 H, H-1), 9.91 (s, 1 H, CHO)C₁₉H₂₁BrO₂, 360.072,

Estra-1,3,5-(10),16-tetraen-17-(1H-benzimidazol-1-yl)-16-formyl-3-ol(VNPP314)

A mixture of compound VNPP312 (0.42 g, 1.16 mmol), benzimidazole (0.41g, 3.48 mmol), and K₂CO₃ (0.48 g, 3.48 mmol) in dry DMF (5 mL) washeated at 80° C. under argon for 5 h. After cooling to room temperature,the reaction mixture was poured onto ice-cold water (100 mL), andresulting solid filtered, washed with water, dried and carried to nextstep without purification. VNPP314 (0.39 g, 84%): mp 255-257° C.

3-(hydroxy)-17-(1H-benzimidazol-1-yl)-estra-1,3,5-(10)-trien (VNPP338(E21))

A solution of VNPP314 (0.39 g, 0.97 mmol) in dry benzonitrile (3 mL) wasrefluxed in the presence of 10% Pd on activated charcoal (0.2 g, i.e.,50% weight of the substrate) for 12 h. Then 0.1 g more catalyst added atinterval of 12 h×2 (all together 0.4 g). After cooling to roomtemperature, the catalyst was removed by filtration through a Celitepad. The filtrate was evaporated, and the residue was purified by FCC[petroleum ether/EtOAc/TEA (7.5:2:0.5) then (1:1:0.1)] to give VNPP338(0.022 g, 6%): mp 110-112° C.; IR (Neat) 1666, 1608, 1493, 1455, 1289,1225, 743; ¹H NMR (500 MHz, CDCl₃) δ 1.02 (s, 3 H, 18-CH₃), 1.48-1.56(m, 3 H), 1.58 (br. s., 7H), 1.70 (d, J=11.14 Hz, 1 H), 1.80-1.89 (m, 1H), 1.90-2.05 (m, 3 H), 2.24-2.33 (m, 1 H), 2.33-2.45 (m, 2 H),2.48-2.56 (m, 1 H), 2.88-2.95 (m, 1 H), 6.02 (br. s., 1 H, 16-H),6.58-6.63 (dd, J=3.0 Hz, 1 H, 4-H), 6.65 (dd, J=8.39, 3.0 Hz, 1 H, 2-H),7.13 (d, J=8.54 Hz, 1 H, 1-H), 7.28-7.35 (m, 2 H, Ar-6¹ and 5¹ Hs),7.49-7.57 (m, 1 H, Ar-7¹ H), 7.79-7.87 (m, 1 H, Ar-4¹ H), 8.00 (s, 1 H,Ar-2¹); ¹³C NMR (500 MHz, CDCl₃) δ 154.6, 147.4, 141.7, 138.0, 132.3,131.7, 128.8, 127.5, 126.2, 123.8, 122.9, 120.1, 115.7, 113.2, 111.4,55.2, 47.9, 44.4, 37.3, 35.1, 30.2, 29.6, 27.4, 26.4, 16.4

Additional Testing:

Luciferase Assay:

Reduction of DHT mediated AR transcriptional activity was performedafter treatment of 10 μM concentration of compounds for 24 h.

Western Blotting:

The ability of various compounds to down-regulate AR protein expressionswas determined by western blotting.

Cell Growth Inhibition (MTT Colorimetric Assay):

Compounds were evaluated on their ability to inhibit LNCaP cell(androgen sensitive AR-mutant), C4-2B and CWR22Rv1 (CRPC cell lines)viability at 1 and 10 μM concentrations by the MTT assy.

Structure Activity Relationship (SAR)

Some nonsteroidal and steroidal agents reduced DHT mediated ARtranscriptional activity in Luciferase assay (FIG. 4). Estrone-3-methylester derivative (VNPP334; Sch. 3) is a potent antiproliferative agentin LNCaP and C4-2B cell line, being more potent than compound VN/124-1(FIGS. 5 and 6).

Of the nonsteroidal agents tested against LNCaP cell line (FIG. 5)methoxy substituted biphenyl with one carbon spacing (VNPP347B; Sch. 1)and naphthalene (VNPP372-2; Sch. 1) derivative displayed more potentanticancer activity in comparison to compound VN/124-1. Whereas,hydroxyl (VNPP356) and nitrile (VNPP358; Sch. 1) substituted biphenylswith one carbon spacing to BzIM group are as potent as compoundVN/124-1.

Estrogen-3-methyl ester derivative (VNPP334; Sch. 3) is more potent thancompound VN/124-1.

From above this limited SAR, it appears that biphenyl derivatives withtwo carbon spacing between BzIM group is detrimental to activity.

Substitution —COOH or 1H-terazole on biphenyl is also detrimental toactivity.

Potent anticancer activity observed with VNPP334, VNPP367 possibly dueto AR down-regulating action is supported by western blot (FIG. 8). Inaddition, equal potent anticancer activity to compound VN/124-1 isdisplayed by VNPP338, VNPP356, which are also potent AR down-regulatingagents.

Two of moderately acting compounds (VNPP341, VNPP358) are ARup-regulators. Their observed anticancer activity may also be due toother yet unknown activities.

MDV3100 and Casodex are not AR down-regulating agents.

Effects of Compounds on Oncogenes.

In the experiments show in FIG. 9, LNCaP and CWR22Rv1 cells were treatedVN/124-1 and analogs at varying time points and effects on significantcancer cells promoting genes investigated at the protein level.

FIGS. 9A and 9C show, in LNCaP cells, Galeterone and analogs (VNPT55 andVNPP414) induced a significant increase in PARP cleavage (signature ofapoptosis) and reduced the expression of CDC25c, Mnk2, Cyclin D1, Cox-2and N-Cadherin.

FIGS. 9B and 9G show in CWR22Rv1 cells Galeterone and analogs (VNPT55and VNPP414) decreased the protein expression of N-Cadherin, AR3,Mnk1/2, MMP-2, Ezh2, Mc1-1(antiapoptotic protein) and also downregulatedthe phosphorylation of eIF4E.

FIGS. 9D, 9E and 9F show CWR22Rv1 were treated with other analogs ofGaleterone and VNPP356, VNPP360, VNPP341, VNPP367, VNPP334 and VNPP338showed activity (AR down-regulation) against full length AR and thesplice variants ARs.

FIG. 9H shows activity (AR down-regulation) of VN-PP433-2b(VNPP-433-6β), VNPP-433-2c (VNPP433-3α (A3)), VNPP433-2d (VNPP433-3β(A1)), and VNPP-334 (A2) against full length AR.

FIG. 10 shows PC3 cells treated with Galeterone, VNPT55 and VNPP414 at 5μM profoundly inhibit migration of PC3 cells after 12 hours.

FIGS. 93A, 9B, 9C & 9D show LNCaP, CWR22Rv1, PC3 and DU145 cellsrespectively were plated and treated with Galeterone, VNPT55,Abiraterone and MDV3100 for duration of 14 to 20 days and colonyformation of the cells determined by staining. As compared toAbiraterone and MDv3100, Galeterone and VNPT55 profoundly inhibitedcolony formation of both AR positive and negative prostate cancer cells.

TABLE 10 GI₅₀ values of VN/124-1 and its analogs GI₅₀ Values (μM)^(a)LNCaP LNCaP Compounds (Low Passage) (High Passage) CWR22Rv1 VN/124-12.45 9.57 4.46 MDV3100 4.85 nd nd VNPT55 0.87 4.06 3.35 VNPP334 (E6) nd6.91 8.74 VNPP414 (A16) 0.87 2.45 3.24 VNPP433-2d (A1) 0.81 1.64 2.54^(a)The GI50 values were determined from dose-response curves (bynonlinear regression analysis using GraphPad Prism) compiled from atleast three independent experiments, SEM <10%, and represents thecompound concentration required to inhibit cell growth by 50%. nd = notdetermined.

The new scaffolds (both steroidal and nonsteroidal) are well tolerated.

3-methylcarboxylate of estrone, methoxy of biphenyl with one carbonspacing and naphthalene derivatives displayed potent biologicalactivity.

Determination of antiandrogenic and CYP17 inhibitory activity of novelagents may delineate their observed anticancer and ARDA activity.

Applicants' invention includes novel compounds and their use onmodulation of the androgen receptor (AR). Applicants have evaluatedtheir effects on AR and other prostate cancer oncogenic targets. Inaddition to modulation of AR (degradation of both full-length AR andsplice variant AR), these novel compounds also degrade MAPK-interactingkinases (Mnk1/2) and modulation eukaryotic translation initiation factor4 (eIF4E). As shown in FIG. 9A-H, 10 and Table 10, the lead compoundsdegrade Mnk1/2, blockeIF4E phosphorylation, inhibit cell growth,colonization, and migration and induce apoptosis. These effects are alsosuperior to some FDA approved prostate cancer drugs.

Androgen receptor associated conditions that may be prevented and/ortreated in the present invention include, but are not limited to,prostate cancer, castration resistant prostate cancer, bladder cancer,pancreatic cancer, hepatocellular carcinoma, benign prostatichyperplasia (BPH), Kennedy's disease, androgenetic alopecia, breastcancer, androgen-insensitive syndrome, and spinal and bulbar muscularatrophy.

GI₅₀Values

Cell Growth Inhibition (MTT Colorimetric Assay):

The cells were seeded in 96-well plates (Corning Costar) at a density of5×10³ cells per well. Cells were allowed to adhere to the plate for 24hours and then treated with various concentrations of compoundsdissolved in 95% EtOH. Cells were treated for 7 days with renewal oftest compound and media on day 4. On the 7th day, medium was renewed andMTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide)(Sigma, St Louis, Mo., USA) solution (0.5 mg MTT per ml of media) wasadded to the medium such that the ratio of MTT: medium was 1:10. Thecells were incubated with MTT for 2 hours. The medium was then aspiratedand DMSO was added to solubilize the violet MTT-formazan product. Theabsorbance at 562 nm was measured by spectrophotometry (Biotek Inc.).The results are shown below.

Steroidal VNPP Compounds

Low Passage High Passage Compound LNCaP LNCaP CWR22Rv1 VN/124-1 3.938.91 MDV-3100 4.85 Abiraterone Acetate VNPP55 4.06 4.46 VNPP334  

6.91 8.74 VNPP397  

9.57 7.52 VNPP412  

8.71 7.50 VNPP414  

0.87 2.45 3.24 VNPP433-6β  

6.61 12.30 VNPP433-3α  

4.04 8.91 9.88 VNPP433-3β  

0.81 1.64 2.54Non-steroidal VNPP Compounds

Low High Passage Passage Compound LNCaP LNCaP CWR22Rv1 VN/124-1 3.938.91 MDV-3100 4.85 Abiraterone Acetate VNPP347B  

4.41 14.00 VNPP358  

3.71 3.50 10.47 VNPP372/2  

5.77 6.09 8.91 VNPP431  

5.23 3.14 4.78Pharmaceutical Composition/Formulation

A pharmaceutical composition, as used herein, refers to a mixture of acompound of the invention with other chemical components, such ascarriers, stabilizers, diluents, dispersing agents, suspending agents,thickening agents, and/or excipients. The pharmaceutical compositionfacilitates administration of the compound to an organism.Pharmaceutical composition containing a compound of the invention can beadministered in therapeutically effective amounts as pharmaceuticalcompositions by any conventional form and route known in the artincluding, but not limited to: intravenous, oral, rectal, aerosol,parenteral, ophthalmic, pulmonary, transdermal, vaginal, optic, nasal,and topical administration. One may administer the compound in a localrather than systemic manner, for example, via injection of the compounddirectly into an organ, often in a depot or sustained releaseformulation. Furthermore, one may administer pharmaceutical compositioncontaining a compound of the invention in a targeted drug deliverysystem, for example, in a liposome coated with organ-specific antibody.The liposomes will be targeted to and taken up selectively by the organ.In addition, the pharmaceutical composition containing a compound of theinvention may be provided in the form of a rapid release formulation, inthe form of an extended release formulation, or in the form of anintermediate release formulation. For oral administration, a compound ofthe invention can be formulated readily by combining the activecompounds with pharmaceutically acceptable carriers or excipients wellknown in the art. Such carriers enable the compounds described herein tobe formulated as tablets, powders, pills, dragees, capsules, liquids,gels, syrups, elixirs, slurries, suspensions and the like, for oralingestion by a patient to be treated.

Pharmaceutical preparations for oral use can be obtained by mixing oneor more solid excipient with one or more of the compounds describedherein, optionally grinding the resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Dragee cores are provided with suitablecoatings. For this purpose, concentrated sugar solutions may be used,which may optionally contain gum arabic, talc, polyvinylpyrrolidone,carbopol gel, polyethylene glycol, and/or titanium dioxide, lacquersolutions, and suitable organic solvents or solvent mixtures. Dyestuffsor pigments may be added to the tablets or dragee coatings foridentification or to characterize different combinations of activecompound doses.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. In someembodiments, the capsule comprises a hard gelatin capsule comprising oneor more of pharmaceutical, bovine, and plant gelatins. In certaininstances, a gelatin is alkaline processed. The push-fit capsules cancontain the active ingredients in admixture with filler such as lactose,binders such as starches, and/or lubricants such as talc or magnesiumstearate and, optionally, stabilizers. In soft capsules, the activecompounds may be dissolved or suspended in suitable liquids, such asfatty oils, liquid paraffin, or liquid polyethylene glycols. Inaddition, stabilizers may be added. All formulations for oraladministration should be in dosages suitable for such administration.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, or gels formulated in conventional manner.Parental injections may involve for bolus injection or continuousinfusion. The pharmaceutical composition of a compound of the inventionmay be in a form suitable for parenteral injection as a sterilesuspensions, solutions or emulsions in oily or aqueous vehicles, and maycontain formulatory agents such as suspending, stabilizing and/ordispersing agents. Pharmaceutical formulations for parenteraladministration include aqueous solutions of the active compounds inwater-soluble form. Additionally, suspensions of the active compoundsmay be prepared as appropriate oily injection suspensions. Suitablelipophilic solvents or vehicles include fatty oils such as sesame oil,or synthetic fatty acid esters, such as ethyl oleate or triglycerides,or liposomes. Aqueous injection suspensions may contain substances whichincrease the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may alsocontain suitable stabilizers or agents which increase the solubility ofthe compounds to allow for the preparation of highly concentratedsolutions. Alternatively, the active ingredient may be in powder formfor constitution with a suitable vehicle, e.g., sterile pyrogen-freewater, before use.

In some embodiments, pharmaceutical formulations are prepared as anamorphous solid dispersion. For example, the pharmaceutical formulationis a spray-dried formulation of the active compound in a polymer matrix.The pharmaceutical formulation may also be prepared by hot meltextrusion.

A compound of the invention can be administered topically and can beformulated into a variety of topically administrable compositions, suchas solutions, suspensions, lotions, gels, pastes, medicated sticks,balms, creams or ointments. Such pharmaceutical compounds can containsolubilizers, stabilizers, tonicity enhancing agents, buffers andpreservatives. Formulations suitable for transdermal administration ofcompounds of the invention may employ transdermal delivery devices andtransdermal delivery patches and can be lipophilic emulsions orbuffered, aqueous solutions, dissolved and/or dispersed in a polymer oran adhesive. Such patches may be constructed for continuous, pulsatile,or on demand delivery of pharmaceutical agents. Still further,transdermal delivery of a compound of the invention can be accomplishedby means of iontophoretic patches and the like. Additionally,transdermal patches can provide controlled delivery of a compound of theinvention. The rate of absorption can be slowed by usingrate-controlling membranes or by trapping the compound within a polymermatrix or gel. Conversely, absorption enhancers can be used to increaseabsorption. An absorption enhancer or carrier can include absorbablepharmaceutically acceptable solvents to assist passage through the skin.For example, transdermal devices are in the form of a bandage comprisinga backing member, a reservoir containing the compound optionally withcarriers, optionally a rate controlling barrier to deliver the compoundto the skin of the host at a controlled and predetermined rate over aprolonged period of time, and means to secure the device to the skin.

For administration by inhalation, a compound of the invention may be ina form as an aerosol, a mist or a powder. Pharmaceutical compositions ofthe invention are conveniently delivered in the form of an aerosol spraypresentation from pressurized packs or a nebuliser, with the use of asuitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol the dosage unitmay be determined by providing a valve to deliver a metered amount.Capsules and cartridges of, such as, by way of example only, gelatin foruse in an inhaler or insufflator may be formulated containing a powdermix of the compound and a suitable powder base such as lactose orstarch.

A compound of the invention may also be formulated in rectalcompositions such as enemas, rectal gels, rectal foams, rectal aerosols,suppositories, jelly suppositories, or retention enemas, containingconventional suppository bases such as cocoa butter or other glycerides,as well as synthetic polymers such as polyvinylpyrrolidone, PEG, and thelike. In suppository forms of the compositions, a low-melting wax suchas, but not limited to, a mixture of fatty acid glycerides, optionallyin combination with cocoa butter is first melted.

In practicing the methods of treatment or use provided herein,therapeutically effective amounts of a compound of the inventionprovided herein are administered in a pharmaceutical composition to amammal having a disease or condition to be treated. In some embodiments,the mammal is a human. A therapeutically effective amount can varywidely depending on the severity of the disease, the age and relativehealth of the subject, the potency of the compound used and otherfactors. The compounds can be used singly or in combination with one ormore therapeutic agents as components of mixtures. Pharmaceuticalcompositions may be formulated in conventional manner using one or morephysiologically acceptable carriers comprising excipients andauxiliaries which facilitate processing of the active compounds intopreparations which can be used pharmaceutically. Proper formulation isdependent upon the route of administration chosen. Any of the well-knowntechniques, carriers, and excipients may be used as suitable and asunderstood in the art. Pharmaceutical compositions comprising a compoundof the invention may be manufactured in a conventional manner, such as,by way of example only, by means of conventional mixing, dissolving,granulating, dragee-making, levigating, emulsifying, encapsulating,entrapping or compression processes.

The pharmaceutical compositions will include at least onepharmaceutically acceptable carrier, diluent or excipient and a compoundof the invention described herein as an active ingredient in free-baseform, or in a pharmaceutically acceptable salt form. In addition, themethods and pharmaceutical compositions described herein include the useof N-oxides, crystalline forms (also known as polymorphs), as well asactive metabolites of these compounds having the same type of activity.

Methods for the preparation of compositions comprising the compoundsdescribed herein include formulating the compounds with one or moreinert, pharmaceutically acceptable excipients or carriers to form asolid, semi-solid or liquid. Solid compositions include, but are notlimited to, powders, tablets, dispersible granules, capsules, cachets,and suppositories. Liquid compositions include solutions in which acompound is dissolved, emulsions comprising a compound, or a solutioncontaining liposomes, micelles, or nanoparticles comprising a compoundas disclosed herein. Semi-solid compositions include, but are notlimited to, gels, suspensions and creams. The compositions may be inliquid solutions or suspensions, solid forms suitable for solution orsuspension in a liquid prior to use, or as emulsions. These compositionsmay also contain minor amounts of nontoxic, auxiliary substances, suchas wetting or emulsifying agents, pH buffering agents, and so forth.

A summary of pharmaceutical compositions described herein may be found,for example, in Remington: The Science and Practice of Pharmacy,Nineteenth Ed (Easton, Pa.: Mack Publishing Company, 1995); Hoover, JohnE., Remington's Pharmaceutical Sciences, Mack Publishing Co., Easton,Pa. 1975; Liberman, H. A. and Lachman, L., Eds., Pharmaceutical DosageForms, Marcel Decker, New York, N.Y., 1980; and Pharmaceutical DosageForms and Drug Delivery Systems, Seventh Ed. (Lippincott Williams &Wilkins1999), herein incorporated by reference in their entirety.

Methods of Administration and Treatment Methods

A compound of the invention can be used in the preparation ofmedicaments for the treatment of diseases or conditions in which steroidhormone nuclear receptor activity contributes to the pathology and/orsymptoms of the disease. In addition, a method for treating any of thediseases or conditions described herein in a subject in need of suchtreatment, involves administration of pharmaceutical compositionscontaining at least one compound of the invention, or a pharmaceuticallyacceptable salt, pharmaceutically acceptable N-oxide, pharmaceuticallyactive metabolite, pharmaceutically acceptable prodrug, orpharmaceutically acceptable solvate thereof, in therapeuticallyeffective amounts to said subject.

The compositions containing the compound(s) described herein can beadministered for prophylactic and/or therapeutic treatments. Intherapeutic applications, the compositions are administered to a patientalready suffering from a disease or condition, in an amount sufficientto cure or at least partially arrest the symptoms of the disease orcondition, or to cure, heal, improve, or ameliorate the conditionitself. Amounts effective for this use will depend on the severity andcourse of the disease or condition, previous therapy, the patient'shealth status, weight, and response to the drugs, and the judgment ofthe treating physician. It is considered well within the skill of theart for one to determine such therapeutically effective amounts byroutine experimentation (including, but not limited to, a doseescalation clinical trial).

In the case wherein the patient's condition does not improve, upon thedoctor's discretion the administration of the compounds may beadministered chronically, that is, for an extended period of time,including throughout the duration of the patient's life in order toameliorate or otherwise control or limit the symptoms of the patient'sdisease or condition. In the case wherein the patient's status doesimprove, upon the doctor's discretion the administration of thecompounds may be given continuously or temporarily suspended for acertain length of time (i.e., a “drug holiday”).

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, the dosage or thefrequency of administration, or both, can be reduced, as a function ofthe symptoms, to a level at which the improved disease or condition isretained. Patients can, however, require intermittent treatment on along-term basis upon any recurrence of symptoms.

In certain instances, it may be appropriate to administertherapeutically effective amounts of at least one of the compoundsdescribed herein (or a pharmaceutically acceptable salts,pharmaceutically acceptable N-oxides, pharmaceutically activemetabolites, pharmaceutically acceptable prodrugs, and pharmaceuticallyacceptable solvates thereof) in combination with another therapeuticagent. By way of example only, if one of the side effects experienced bya patient upon receiving one of the compounds herein is inflammation,then it may be appropriate to administer an anti-inflammatory agent incombination with the initial therapeutic agent. Or, by way of exampleonly, the therapeutic effectiveness of one of the compounds describedherein may be enhanced by administration of an adjuvant (i.e., by itselfthe adjuvant may only have minimal therapeutic benefit, but incombination with another therapeutic agent, the overall therapeuticbenefit to the patient is enhanced). Or, by way of example only, thebenefit of experienced by a patient may be increased by administeringone of the compounds described herein with another therapeutic agent(which also includes a therapeutic regimen) that also has therapeuticbenefit. In any case, regardless of the disease or condition beingtreated, the overall benefit experienced by the patient may simply beadditive of the two therapeutic agents or the patient may experience asynergistic benefit. Where the compounds described herein areadministered in conjunction with other therapies, dosages of theco-administered compounds will of course vary depending on the type ofco-drug employed, on the specific drug employed, on the disease orcondition being treated and so forth. In addition, when co-administeredwith one or more biologically active agents, the compound providedherein may be administered either simultaneously with the biologicallyactive agent(s), or sequentially. If administered sequentially, theattending physician will decide on the appropriate sequence ofadministering protein in combination with the biologically activeagent(s).

In any case, the multiple therapeutic agents (one of which is one of thecompounds described herein) may be administered in any order or evensimultaneously. If simultaneously, the multiple therapeutic agents maybe provided in a single, unified form, or in multiple forms (by way ofexample only, either as a single pill or as two separate pills). One ofthe therapeutic agents may be given in multiple doses, or both may begiven as multiple doses. If not simultaneous, the timing between themultiple doses may vary from more than zero weeks to less than fourweeks. In addition, the combination methods, compositions andformulations are not to be limited to the use of only two agents.Multiple therapeutic combinations are envisioned.

In addition, a compound of the invention may also be used in combinationwith procedures that may provide additional or synergistic benefit tothe patient. By way of example only, patients are expected to findtherapeutic and/or prophylactic benefit in the methods described herein,wherein pharmaceutical composition of the invention and/or combinationswith other therapeutics are combined with genetic testing to determinewhether that individual is a carrier of a mutant gene that is known tobe correlated with certain diseases or conditions. A compound of theinvention and combination therapies can be administered before, duringor after the occurrence of a disease or condition, and the timing ofadministering the composition containing a compound can vary. Thus, forexample, the compounds can be used as a prophylactic and can beadministered continuously to subjects with a propensity to conditions ordiseases in order to prevent the occurrence of the disease or condition.The compounds and compositions can be administered to a subject duringor as soon as possible after the onset of the symptoms. Theadministration of the compounds can be initiated within the first 48hours of the onset of the symptoms, preferably within the first 48 hoursof the onset of the symptoms, more preferably within the first 6 hoursof the onset of the symptoms, and most preferably within 3 hours of theonset of the symptoms. The initial administration can be via any routepractical, such as, for example, an intravenous injection, a bolusinjection, infusion over 5 minutes to about 5 hours, a pill, a capsule,transdermal patch, buccal delivery, and the like, or combinationthereof. a compound is preferably administered as soon as is practicableafter the onset of a disease or condition is detected or suspected, andfor a length of time necessary for the treatment of the disease, suchas, for example, from about 1 month to about 3 months. The length oftreatment can vary for each subject, and the length can be determinedusing the known criteria. For example, the compound or a formulationcontaining the compound can be administered for at least 2 weeks,preferably about 1 month to about 3 years, and in some embodiments fromabout 1 month to about 10 years.

The pharmaceutical composition described herein may be in unit dosageforms suitable for single administration of precise dosages. In unitdosage form, the formulation is divided into unit doses containingappropriate quantities of one or more compound. The unit dosage may bein the form of a package containing discrete quantities of theformulation. Non-limiting examples are packaged tablets or capsules, andpowders in vials or ampoules. Aqueous suspension compositions can bepackaged in single-dose non-reclosable containers. Alternatively,multiple-dose reclosable containers can be used, in which case it istypical to include a preservative in the composition. By way of exampleonly, formulations for parenteral injection may be presented in unitdosage form, which include, but are not limited to ampoules, or inmulti-dose containers, with an added preservative.

The invention claimed is:
 1. A method of treating cancer comprisingadministering an effective amount of a compound selected from the groupconsisting of3β-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP433-3β (A1)),3β-(pyridine-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP414(A16)), 6-(3-benzimidazolylphenyl)naphthalen-2-ol(VNPP372/2(Na1)), and N-(3-benzimidazolylphenyl)[(4-methoxyphenyl)sulphonyl]amine (VNPP431(SulAmd2)) to a patient in need thereof, whereinsaid cancer is prostate cancer or castration resistant human prostatecancer.
 2. The method of claim 1, wherein said cancer is prostatecancer.
 3. The method of claim 1, wherein said cancer is castrationresistant human prostate cancer.
 4. A method of degrading a full lengthandrogen receptor or a splice variant androgen receptor, the methodcomprising administering an effective amount of a compound selected fromthe group consisting of3β-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP433-3β (A1)),3β-(pyridin-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP414(A16), 6-(3-benzimidazolylphenyl)naphthalen-2-ol(VNPP372/2(Na1)), and N-(3-benzimidazolylphenyl)[(4-methoxyphenyl)sulphonyl]amine (VNPP431(SulAmd2)) to a patient in need thereof.
 5. Amethod of inhibiting proliferation of androgen sensitive cell linecomprising administering an effective amount of a compound selected fromthe group consisting of3β-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP433-3β (Al)),3β-(pyridin-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP414(A16), 6-(3-benzimidazolylphenyl)naphthalen-2-ol (VNPP372/2(Na1)), and N-(3-benzimidazolylphenyl)[(4-methoxyphenyl) sulphonyl]amine(VNPP431(SulAmd2)) to a patient in need thereof.
 6. A method of treatingcancer, comprising administering an effective amount of a compoundselected from the group consisting of3β-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP433-3β (A1)), and3β-(pyridin-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP414 (A16)) to a patient in need thereof, wherein the cancer is atleast one selected from the group consisting of prostate cancer andcastration resistant prostate cancer.
 7. The method of treating canceraccording to claim 6, wherein the compound is3,β-(1H-imidazol-1-yl)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP433-3β (A1)).
 8. The method of treating cancer according to claim6, wherein the compound is3β-(pyridin-4-ylmethoxy)-17-(1H-benzimidazol-1-yl)androsta-5,16-diene(VNPP414 (A16)).
 9. The method of treating cancer according to claim 6,wherein the cancer is prostate cancer.
 10. The method of treating canceraccording to claim 6, wherein the cancer is castration resistantprostate cancer.